ReviewArticle Regulation of Breast Cancer and Bone ... · 370 DiseaseMarkers miRNA gene Pol2/3 RISC...
19
Hindawi Publishing Corporation Disease Markers Volume 35 (2013), Issue 5, Pages 369–387 http://dx.doi.org/10.1155/2013/451248 Review Article Regulation of Breast Cancer and Bone Metastasis by MicroRNAs S. Vimalraj, P. J. Miranda, B. Ramyakrishna, and N. Selvamurugan Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur Tamil Nadu, 603 203, India Correspondence should be addressed to N. Selvamurugan; [email protected] Received 17 June 2013; Revised 17 August 2013; Accepted 27 August 2013 Academic Editor: Benoit Dugue Copyright © 2013 S. Vimalraj et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Breast cancer progression including bone metastasis is a complex process involving numerous changes in gene expression and function. MicroRNAs (miRNAs) are small endogenous noncoding RNAs that regulate gene expression by targeting protein-coding mRNAs posttranscriptionally, oſten affecting a number of gene targets simultaneously. Alteration in expression of miRNAs is common in human breast cancer, possessing with either oncogenic or tumor suppressive activity. e expression and the functional role of several miRNAs (miR-206, miR-31, miR-27a/b, miR-21, miR-92a, miR-205, miR-125a/b, miR-10b, miR-155, miR-146a/b, miR- 335, miR-204, miR-211, miR-7, miR-22, miR-126, and miR-17) in breast cancer has been identified. In this review we summarize the experimentally validated targets of up- and downregulated miRNAs and their regulation in breast cancer and bone metastasis for diagnostic and therapeutic purposes. 1. Introduction Breast cancer is a malignant breast neoplasm originating from breast tissues, and its advanced form tends to metas- tasize into bone. It comprises 10.4% of all cancer incidences among women, and it is the most common type of nonskin cancer in women. Breast cancer is about 100 times more common in women than in men although males tend to have poorer outcomes due to delays in diagnosis [1–3]. Breast cancer metastasizes into bone is the process of spreading of the advanced form of breast tumor cells into bone. us the interactions between breast tumor cells and bone cells lead to impede the bone remodeling, in specific breast tumor which is recorded to predominantly increase the osteoclastic activity [4]. e radiation and chemotherapy are the potential and effective for cancer treatment, but they lead to side effects by destroying noncancerous cells or healthy cells, and this type of heavy toxic burden impedes the immune system; hence the patient would be susceptible to other infections. In addition, the continuous treatment of radiation and chemotherapy results in less effective destroying cancer cells because of resistance gained [5, 6]. ere is an urge to bring out novel technique(s) for facilitating the diagnostic and therapeutic approaches for cancer treatment. 1.1. Molecular Diagnostics and erapies for Breast Cancer. e traditional clinical approach to treat breast cancer involves a combination of existing surgical-, chemical-, and radiation-based therapies. Surgical options include lumpec- tomy, quadrantectomy, mastectomy, and modified radical mastectomy. ese procedures are sometimes followed by adjuvant therapy depending on the body conditions of patients [7]. Hormonal therapies include selective estro- gen receptor modulators (SERMs), such as tamoxifen, and aromatase inhibitors, such as anastrozole. Chemotherapy includes traditional chemotherapies as well as specific drugs such as trastuzumab, a monoclonal antibody to the HER2/ neu receptor. ere may be possibility of toxicities or unwanted side effects associated with their administration that negatively affect the patient [8, 9]. Molecular diagnostic tests deal with personalized diag- nostic information and allow specific treatment plans con- fining resistance, non response, and toxicity. e analysis of miRNAs may identify their role in decision making process for diagnostic and therapeutic approaches. Several RNA- based molecular diagnostic tools such as microarrays or quantitative reverse transcription (qRT)-PCR analysis focus on gene expression [9, 10]. Recently several studies indicate the significant differential expression and functional role of
Transcript of ReviewArticle Regulation of Breast Cancer and Bone ... · 370 DiseaseMarkers miRNA gene Pol2/3 RISC...
Hindawi Publishing CorporationDisease MarkersVolume 35 (2013) Issue 5 Pages 369ndash387httpdxdoiorg1011552013451248
Review ArticleRegulation of Breast Cancer and Bone Metastasis by MicroRNAs
S Vimalraj P J Miranda B Ramyakrishna and N Selvamurugan
Department of Biotechnology School of Bioengineering SRM University Kattankulathur Tamil Nadu 603 203 India
Correspondence should be addressed to N Selvamurugan selvamn2yahoocom
Received 17 June 2013 Revised 17 August 2013 Accepted 27 August 2013
Academic Editor Benoit Dugue
Copyright copy 2013 S Vimalraj et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited
Breast cancer progression including bone metastasis is a complex process involving numerous changes in gene expression andfunction MicroRNAs (miRNAs) are small endogenous noncoding RNAs that regulate gene expression by targeting protein-codingmRNAs posttranscriptionally often affecting a number of gene targets simultaneously Alteration in expression of miRNAs iscommon in human breast cancer possessing with either oncogenic or tumor suppressive activityThe expression and the functionalrole of several miRNAs (miR-206 miR-31 miR-27ab miR-21 miR-92a miR-205 miR-125ab miR-10b miR-155 miR-146ab miR-335 miR-204 miR-211 miR-7 miR-22 miR-126 and miR-17) in breast cancer has been identified In this review we summarize theexperimentally validated targets of up- and downregulated miRNAs and their regulation in breast cancer and bone metastasis fordiagnostic and therapeutic purposes
1 Introduction
Breast cancer is a malignant breast neoplasm originatingfrom breast tissues and its advanced form tends to metas-tasize into bone It comprises 104 of all cancer incidencesamong women and it is the most common type of nonskincancer in women Breast cancer is about 100 times morecommon in women than in men although males tend tohave poorer outcomes due to delays in diagnosis [1ndash3] Breastcancer metastasizes into bone is the process of spreading ofthe advanced form of breast tumor cells into bone Thus theinteractions between breast tumor cells and bone cells lead toimpede the bone remodeling in specific breast tumor whichis recorded to predominantly increase the osteoclastic activity[4] The radiation and chemotherapy are the potential andeffective for cancer treatment but they lead to side effects bydestroying noncancerous cells or healthy cells and this typeof heavy toxic burden impedes the immune system hence thepatient would be susceptible to other infections In additionthe continuous treatment of radiation and chemotherapyresults in less effective destroying cancer cells because ofresistance gained [5 6] There is an urge to bring out noveltechnique(s) for facilitating the diagnostic and therapeuticapproaches for cancer treatment
11 Molecular Diagnostics and Therapies for Breast CancerThe traditional clinical approach to treat breast cancerinvolves a combination of existing surgical- chemical- andradiation-based therapies Surgical options include lumpec-tomy quadrantectomy mastectomy and modified radicalmastectomy These procedures are sometimes followed byadjuvant therapy depending on the body conditions ofpatients [7] Hormonal therapies include selective estro-gen receptor modulators (SERMs) such as tamoxifen andaromatase inhibitors such as anastrozole Chemotherapyincludes traditional chemotherapies as well as specific drugssuch as trastuzumab a monoclonal antibody to the HER2neu receptor There may be possibility of toxicities orunwanted side effects associated with their administrationthat negatively affect the patient [8 9]
Molecular diagnostic tests deal with personalized diag-nostic information and allow specific treatment plans con-fining resistance non response and toxicity The analysis ofmiRNAs may identify their role in decision making processfor diagnostic and therapeutic approaches Several RNA-based molecular diagnostic tools such as microarrays orquantitative reverse transcription (qRT)-PCR analysis focuson gene expression [9 10] Recently several studies indicatethe significant differential expression and functional role of
370 Disease Markers
miRNA gene
Pol23
RISCloading
complex
Degradation
GW182
Ago
RISCcomplex
Microprocessorcomplex
Drosha
DGCR8
DicerTRBP
Translationrepression
mRNA
Exportin 5
Inhibition of mRNA deadenylationtranslation initiation
(GW182 TRBPFMPR1 Ago Dicer
PABP eIF6)
translation elongation(GW182 TRBP
FMPR1 Ago DicerPABP)
(GW182 TRBP AgoFMPR1 Dicer
PABP CCR4 CAFPan23)
Inhibition of
AAA3 998400
5 998400
Ago
Pre-miRNA
Pri-miRNA
Pre-miRNA
Duplex miRNA
Figure 1 Biogenesis and function of microRNAs RNA polymerase 23 binds to the promoter region of specific DNA sequence and forms ahair pin structure of the pri-miRNA DGCR8 (Pasha) associates with protein Drosha which process pri-miRNA to pre-miRNA (by cleavingnucleotides from hair pin) Pre-miRNA is exported into cytoplasmwith shuttle protein exportin Inmammals the pre-miRNA is loaded ontoa multiprotein complex consisting minimally of Dicer Tar RNA Binding Protein (TRBP) and Ago2 The RNase III enzyme Dicer processesthe precursor by cleaving the stem loop to produce thematuremiRNA (canonical pathway) [11ndash13]The precursor can be loaded directly ontoAgo2 protein and this Argonaute cleaves the precursor RNA of this miRNA [13] The 51015840 nucleotide of the miRNA guide strand especially 51015840U functions as an additive anchor and helps Ago proteins to load mismatch-containing miRNAmiRNA duplexes [14] Of the two strandsthe guide strand is integrated with RISC forming miRNA-RISC complex and other strand is degraded [13ndash16] RISC is involved in inhibitionof translation initiation inhibition of translation elongation or mRNA deadenylation [15ndash17]
miRNAs in breast cancer bonemetastases and other cancerssuggesting that miRNAs could be a valuable biomarker forcancers However the regulatory mechanisms of miRNAs inbreast cancer and bone metastasis remain unclear Here wehave systematically summarized the experimentally validatedtargets of up- and downregulated miRNAs along with theirregulation in breast cancer and bonemetastasis for diagnosticand therapeutic purposes
12 MicroRNAs Synthesis MicroRNAs (miRNAs) are a classof tiny noncoding endogenous RNA molecules only 18ndash25nucleotides long These small molecules have been shownto play critical regulatory roles in a wide range of bio-logical and pathological processes miRNAs may regulatecellular gene expression at the posttranscriptional level bysuppressing translation of protein coding genes or cleavingtarget mRNAs to induce their degradation through imperfectpairing with target mRNAs of protein coding genes at 31015840UTR(untranslated region) The interacting region of 51015840 end ofmiRNA nucleotides (2 to 8 nt) is called seed sequence [18ndash20] However few reports emphasized that miRNAs can alsotarget at 51015840UTRs and protein coding regions of mRNA [1819 21 22] According to miRBase 2578 mature and 1872precursor forms of miRNAs have been identified in human(httpwwwmirbaseorg) MicroRNAs are transcribed with
RNA polymerase II (Figure 1) The RNA polymerase bindsto the promoter region of specific DNA sequence and formsa hair pin structure of the pri-miRNA The transcript thusobtained is methylated at 51015840 region and polyadenylatedat 31015840 region which undergoes posttranscriptional splicingresulting in pri-miRNA If the stem loop is found in 31015840UTRthe transcript serves as pri-miRNA [11 18] The pri-miRNA(poly or monocystronic) has about 1 kb nucleotides in lengthand its double stranded region of hair pin structure isrecognized by nuclear protein DiGeorge syndrome criticalregion 8 (DGCR8or ldquoPashardquo in invertebrates) [11 12] DGCR8associates with protein Drosha which process pri-miRNAto pre-miRNA (by cleaving nucleotides from hair pin) Pre-miRNAs (sim70 nt) have 31015840 overhang due to cleavage by RNApol II Pre-miRNA is exported into cytoplasm with shuttleprotein exportin This is an energy coupled transport withGTP bound Ran protein a nuclear transport receptor [13]
In cytoplasm the Dicer an RNase III enzyme cleaves pre-miRNA to give double strandedmiRNAduplex [14] Inmam-mals the pre-miRNA is loaded onto a multiprotein complexconsisting minimally of Dicer Tar RNA Binding Protein(TRBP) and Ago2 The Dicer processes the precursor bycleaving the stem loop to produce the mature miRNA Ago2binds with mature miRNA to form the RISC (RNA inducedsilencing complex) complex to target specific mRNA This iscanonical miRNA biogenesis (Figure 1) miRNA biogenesis
Disease Markers 371
Sample typeNormalTumor
T20
T22
T43
T17
T18
T25T4T31
T27
T35
T24
T51T5T38T2T42
T13
T29
T36
T40
T26T1T14
T37
N1
N4
T34
T49
T16
T46
T28
T21
T30T9T6T45
T48
T12
N2
T50
T15
N3
T23T8T32
T33
T47
T41
T39T3T19
T10T7T44
T11
Sample type
hsa-miR-27ahsa-miR-21hsa-miR-27bhsa-miR-31hsa-miR-205hsa-miR-206hsa-miR-125bhsa-miR-125a-5phsa-miR-10bhsa-miR-155
hsa-miR-146ahsa-miR-146b-5p
Figure 2 Heat map of differential expression of miRNAs in human normal and tumor breast cells Differential expression of miR-206 miR-31 miR-27a miR-21 miR-27b miR-205 miR-125b miR-125a-5p miR-10b miR-155 miR-146b-5p and miR-146a was identified in normalbreast and breast tumor cells by miRNA body map along with the hierarchical cluster analysis Expression of miRNAs is represented as blue(downregulated) red (upregulated) and white colors (no significant change or absence of data) Here T represents breast tumor cells and Nrepresents breast normal cells
can also depend on Ago2 protein instead of Dicer In caseof miR-451 the precursor is loaded directly onto Ago2 andthis protein cleaves the pre-miRNA The mature miR-451 isbound by Ago2 to form the RISC (RNA induced silencingcomplex) that targets erythropoietic-specific mRNAs [15]The 51015840 phosphate of the miRNA strand is essential for duplexloading into Ago and the preferred 51015840 nucleotide (uridine)of the miRNA strand (guide strand) and the base pairingstatus in the seed region and the middle of the 31015840 regionfunction as additive anchors to Ago [16] Of the two strandsthe guide strand is integratedwithRISC andother strand thatis passenger stand is degraded due to its instability [16 17]The miRNARISC complex attaches to the messenger RNA(mRNA) in one of the two ways (1) When the sequencesare perfectly complementary the miRNARISC complexbinds tightly to the mRNA and via the enzyme Ago2 themRNA is degraded [23]The poly A binding protein (PABP-)dependent poly(A) nuclease 2 (Pan2-) Pan3 deadenylases areinvolved in mRNA deadenylation that could lead to mRNAdegradation [16 18 23] Poly A seems to give stability to themRNA (2) More commonly when the sequences are imper-fectly complementary the miRNARISC complex binds andinhibits translation of the mRNA without degradation Thefinal outcome of either of these pathways is a decrease in theprotein level of the target gene The question of whether amiRNA induces translation inhibition ormRNA degradationwould depend on the level of individual miRNA specifictargets and cell backgrounds [18 19] (Figure 1)
2 miRNAs Expression
MicroRNAs play an important role in normal functioningof the cells Deregulation of miRNAs has been associated
with several disease conditions [19] Most of the miRNAsare either upregulated or downregulated thereby acting asoncogenes or tumor suppressor genes by regulating their tar-get genes To evaluate the differential expression of preferredmiRNAs between normal and breast cancer cells we per-formed miRNA body map analysis (httpwwwmirnabody-maporg) based on database instructions [24] This databaseholds expression of miRNAs from normal and diseasedtissues by RT-qPCR analysis and the data allows the analysisof differential expression of miRNAs between specific tissuesor samples The heat map provided here emphasizes thedifferential expression of miR-206 miR-31 miR-27a miR-21miR-27b miR-205 miR-125b miR-125a-5p miR-10b miR-155 miR-146b-5p and miR-146a in 51 breast cancer samplesand 4 normal breast samples of human (Figure 2) Theheat map based on color (blue down regulation and redup regulation) and cluster clearly depicts the differentialexpression of preferred miRNAs between normal and tumorbreast cellsThe up- or downregulatedmiRNAs are correlatedwith the expression of their targeted genes which couldprovide valuable physiological and pathological informationabout their systems [18ndash20]
21 Oncogenic miRNAs
211 miR-21 In breast cancer miR-21 was found to beoverexpressed [10 25] The proteomic analysis revealed thatTPM1 (Tropomysin) expression is reduced in tumor cellsand it is a target gene of miR-21 [25] TPM1 is a tumorsuppressor gene which regulates microfilament organizationand anchorage-independent growth [26] It was found thatthere is a putative miR-21 binding site at the 31015840UTR of TPM1variants When TPM1 was overexpressed in breast cancer
372 Disease Markers
(MCF-7) cells the anchorage-independent growth was sup-pressed [25] A tumor suppressor gene phosphatase andtensin homolog (PTEN) is also targeted by miR-21 [27] Up-regulation of miR-21 limits the activity of PTEN and PTENis known to limit the activity of PI3K pathway [28] Anothertumor suppressor gene product PDCD4 (programmed celldeath) was originally characterized as an inhibitor of cellulartransformation in a mouse cell culture model [29] Studieshave shown that the levels of PDCD4 mRNA as well asprotein were upregulated by miR-21 inhibition indicatingthat PDCD4 is an important functional target for miR-21[30 31] In all-transretinoic acid (ATRA) treatment miR-21 was selectively induced in ER-120572 positive cells and not inER-120572 negative cells Based on differential expression of genesregulated by ATRA in ER-120572 positive and ER-120572 negative cellsthree more direct miR-21 targets (proinflammatory cytokine(IL1B) adhesion molecule (ICAM-1 and PLAT) tissue-typeplasminogen activator (tPA)) have been predicted [32] miR-21 downregulatedMSH2 and SMAD7 in TGF-120573 pathway andcaused breast cancer progression and upregulated humanepidermal growth factor receptor 2 and thus it acted asoncogene [33 34] miR-21 with numerous gene targets inbreast cancer progression is positively regulated by a proteinnamed nucleolin (NCL) a major nucleolar protein at post-transcriptional level Overall it is emphasized that miR-21 isone of the important miRNAs involved in the regulation ofcancer cells [35 36]
212 miR-10b In metastatic breast cancer cell lines miR-10b was upregulated after activation of twist transcriptionfactor and its overexpression increased with tumor size andinvasiveness It appeared that miR-10b inhibits translationof homeobox D10 (HOXD10) resulting in induction of theprometastatic gene product RHOC [37] This RHOC inturn favors cell migration and invasion The initiation ofmetastasis in nonmetastatic cancer with overexpression ofmiR-10b in an in vivo experiment proved its role in themetastasis of breast cancer Interestingly it was identified thatmiR-10b was down-regulated in breast cancer cells comparedto normal breast cells providing evidence that miR-10b doesnot play any role during initial tumor development [38] andproliferation [39] In contrast to its oncogenic role it wasalso identified that miR-10b targeted various oncogenic genessuch as FLT1 BDNF and SHC1 a signal transduction factor[10] The targets of miR-10b identified are BUB1 PLK1 andCCNA2 [40] miR-10b also targets syndecan-1 and promotesbreast cancer metastasis and invasiveness [41]
213 miR-125 Data reported that miR-125 is likely to downregulate genes which promote tumorigenesis Overexpres-sion of miR-125a or mir-125b in an erb2-dependent cancercell line (skbr3) suppressed her2 and her3 transcript andprotein levels which decreased cell motility and invasiveness[42] miR-125a and miR-125b were significantly downregu-lated in her2-positive breast cancers Computation analysisconfirmed target sites at the 31015840UTR regions of her2 and her3for these miRNAs [43] The role of miR-125 as an oncogene
has been validated by its role in suppressing various tumorsuppressor genes including p53 gene [44]
214 miR-155 miR-155 has been shown as over-expressed inbreast cancer and further it is upregulated in normal mousemammary gland epithelial cells (NMuMG cells) by the TGF-120573Smad4 pathway and it mediates TGF-120573-induced EMT andcell invasion [10 45 46] The ectopic expression of miR-155in NMuMG cells disrupted proper tight junction formationand promoted cellmigration and invasion Conversely antag-onizing miR-155 in NMuMG cells reduced the occurrenceof TGF-120573-induced EMT and cell migration and invasionmiR-155 directly inhibits the expression of RhoA a gene thatregulates many cellular processes including cell adhesionmotility and polarity and is an important modulator ofcell junction formation and stability It is speculated thatTGF-120573Smad4 regulates expression of miR-155 resulting indown regulation of RhoA protein expression to drive EMTprogression In another study it was shown that miR-155 isassociated with cancer invasiveness in human primary breastcarcinoma that is miR-155 is highly expressed in invasivetumors but not in noninvasive cancer tissues [45 46]
Ectopic expression ofmiR-155 induces cell survival but itsknockdown leads cell to apoptosis and enhances chemosen-sitivity FOXO3a has been identified as direct target formiR-155 Inverse correlation of FOXO3a and miR-155 hasbeen observed in breast cancer cell lines and tumors miR-155 directly interacts with 31015840UTR of FOXO3a and blocksits translation Hence FOXO3a is negatively regulated bymiR-155 and FOXO3a mediates miR-155 function in thecontrol of breast cancer cell survival and growth [47]Further the phenomenon of up regulation of hexokinase2 (hk2) which enhances the metabolic activity of breastcancer cells by miR-155 makes it as a potential candidate foranticancer therapy miR-155 first activates signal transducerand activator of transcription 3 (STAT3) a transcriptionalactivator for hk2 and secondly target CEBP120573 [48] It hasbeen shown that tumor-associated circulating miRs (miR-10bmiR-34amiR-141 andmiR-155) are elevated in the bloodof breast cancer patients and hence there would be feasibilityand clinical utility of circulating miRNAs as biomarkersfor the detection and staging of breast cancer [49] It wasevident that miR-155 inhibited BMP2- BMP6- and BMP7and induced ID3 expression by targeting SMAD1 SMAD5HIVEP2 CEBPB RUNX2 and MYO10 components of theBMP signaling cascade [50] Recent findings indicate thatmiR-155 contributes to cell proliferation by down regulationof TP53INP1 in estradiol promoted breast cancer cells [51]miR-155 also downregulated tumor suppressor genes such asp53 [52] CDC73 [53] and VHL [54] in breast cancer cells InTable 1 the detailed information about oncogenic miRNAstheir target genes and their regulation during breast cancerprogression is provided
22 Tumor Suppressor miRNAs
221 miR-206 A differential expression of miR-206 wasobserved in cancer and normal tissues miR-206 targets
Disease Markers 373
Table1Targetso
foncogenicmiRNAs
andtheirroleincancer
cells
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-21
TPM1
WB
LAmiR-21d
ownregulates
TPM1inbreastcancer
andleadstoanchorage-independ
entg
rowth
[25]
E2F1
WB
RT-PCR
miR-21regulates
E2F1
byTG
F-120573sig
nalin
gpathway
[55]
TGFB
R2WB
RT-PCR
miR-21regulates
TGFB
R2by
TGF-120573sig
nalin
gpathway
[55]
TIMP3
WB
RT-PCR
TIMP3
isaM
MPinhibitormiR-21sup
pressesT
IMP3
byprom
otinginvasiv
enesso
fcancerc
ells
[56]
SERP
INB5
WB
LADow
nregulationof
PDCD
4andmaspin(SER
PINB5
)bymiR-21causestum
orogenesis
[57]
PDCD
4WB
RT-PCR
LA
miR-21d
ecreases
PDCD
4proteinam
ountsa
ndincreasesinvasion
[58]
FASCD
K6P
DCD
4CD
KN1A
FAM3C
HIPK3
PR
RG4AC
TA2BT
G2
BMPR
2SE
SN1IL6R
SOCS
5GLC
CI1APA
F1
SLC1
6A10SGK3
RP2
CFL
2
RT-PCR
M
Inhibitio
nof
miR-21inMCF
-7breastcancer
cells
show
sreduced
expressio
nof
p53tumor
supp
ressor
protein
FASCD
K6C
DKN
1AFAM3C
HIPK3
PRR
G4AC
TA2BT
G2BM
PR2
SESN
1IL6R
andtumor
supp
ressor
proteinProgrammed
CellD
eath
4(PDCD
4)area
lsodo
wnregulated
bymiR-21
[30]
FASTIMP3
WB
LADow
nregulationof
miR-21increases
FasligandandTIMP3
expressio
ntherebyitisinvolved
intheu
pregulationof
apop
tosis
[59]
CDC2
5AWB
RT-PCR
LA
miR-21sup
pressesC
dc25Aexpressio
nac
ellcycleregulator
[60]
RECK
RT-PCR
miR-21sup
pressesR
ECKandinhibitorsof
MMPs
andthus
prom
otes
invasiv
enesso
fcancerc
ells
[56]
MTA
PLA
Apop
totic
functio
nof
MTA
Pissupp
ressed
bymiR-21
[61]
RECK
WB
RT-PCR
LA
RECK
regu
latingthem
embrane-anchored
protease
isdo
wnregu
lated
bymiR-21
[62]
PDCD
4WB
LAmiR-21p
ositivelyregulates
PDCD
4throug
hinteractionwith
ovarianste
roidsa
ndTG
F-120573
signalin
gpathway
[55]
BTG2
EGFPW
BFluo
rescence
Intensity
Expressio
nof
miR-21leads
tolowe
rlevelof
BTG2which
isac
ellcycleregu
latora
ndtumor
supp
ressorItcauseslosso
fcon
trolonG1toSph
asetransition
[63]
PTEN
LAmiR-21regulates
MMP-2expressio
nin
cardiacfi
brob
lasts
oftheinfarctzone
viaP
TENpathway
[64]
BTG2
WB
RT-PCR
LA
miR-21d
irectlyinhibitsBT
G2which
isac
ellcycleinhibitorleading
toun
controlledDNA
synthesis
byactiv
ationof
forkhead
boxM1
[65]
PELI1
RT-PCR
LA
miR-21o
verexpressioninhibitsPeli1
viak
appaBsig
nalling
[66]
PDCD
4WB
Deregulationof
miR-21b
yvinylcarbam
ateind
uces
mou
selung
tumorigenesisby
downregulating
PDCD
4[67]
BIG-H
3LA
miR-21targetsbig-h3Itcou
ldbe
indu
cedby
TGF-120573anditregu
latesT
GF-120573sig
nalling
positively
andnegativ
ely
[68]
BIM
WB
LABima
apop
totic
activ
atorisc
ollectively
targeted
bymiR-21miR-24andmiR-221
andits
actio
nis
supp
ressed
[69]
HER
2WB
RT-PCR
miR-21u
pregulates
human
epidermalgrow
thfactor
receptor
2andactsas
oncogene
[34]
MSH
2SM
AD7
WB
RT-PCR
LA
miR-21d
ownregulates
MSH
2andSM
AD7in
TGF-120573pathway
andcauses
breastcancer
progression
[33]
374 Disease Markers
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-10b
HOXD
10WB
LAOverexpressionof
miR-10b
indu
cesrob
ustinvasionandmetastasis
bydo
wnregu
latin
gtransla
tionof
homeobo
xD10
[37]
KLF4
WB
RT-PCR
LA
miR-10b
redu
cese
ndogenou
sKLF
4protein
atum
orsupp
ressor
[70]
SFRS
1WB
RT-PCR
LA
miR-10b
supp
resses
SFRS
1(SR
-family
splicingfactor)a
ndcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
SFRS
10LA
miR-10b
supp
resses
SFRS
10andcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
miR-125a-5p
ERBB
2WB
RT-PCR
LA
Expressio
nof
either
miR-125ao
rmiR-125bresults
insupp
ressionof
ERBB
2andER
BB3
(oncogenefam
ily)a
ndexhibitsredu
cedmigratio
nandinvasio
ncapacitie
s[72]
TP53
WB
RT-PCR
LA
miR-125binhibitsp53gene
transla
tionally
andtranscrip
tionally
andtheisomer
ofmiRNA-
125
(miRNA-
125a)translationally
arrests
mRN
Aof
thep
53a
tumor
supp
ressor
gene
[44]
HuR
WB
RT-PCR
LA
HuR
isas
tressindu
cedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
Expressio
nof
miR-125as
ignificantly
redu
cesH
uR
[73]
ARID3B
WB
ARID3B
isoverexpressedin
human
ovariancancerO
verexpressionof
miR-125aind
uces
conversio
nof
high
lyinvasiv
eovaria
ncancer
cells
from
amesenchym
alto
anepith
elial
morph
ologyandsupp
resses
cancer
bynegativ
elyregu
latin
gEM
T[74]
miR-125b
BAK1
WB
RT-PCR
LAM
Overexpressionof
miR-125bstimulates
androgen-in
depend
entg
rowth
ofprostatecancer
bydo
wnregu
latin
gBa
k1(proapop
totic
regu
lator)
[75]
CDK6
CDC2
5AWB
miR-125boverexpressio
ndecreasesC
DK6
andCD
C25A
expressio
nItregu
latesp
roliferationof
U251g
liomas
tem
cells
throug
hcellcycle
regu
latedproteins
CDK6
andCD
C25A
[76]
SMO
WB
RT-PCR
LA
Dow
nregulationof
miR-125ballowsh
ighlevelsof
Hedgeho
g-depend
entgenee
xpressionleading
totumor
cellproliferatio
nby
allowingHedgeho
gsig
nalling
[77]
ST18
WB
BranchedL
AD
NA
Prob
eAssay
ST18
isdo
wnregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
DICER
1WB
LAB
ranchedDNA
Prob
eAssay
DICER
1isd
ownregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
TP53IN
P1LA
miR-125bdirectlyrepressesT
P53INP1anapop
tosis
regulator
inp53pathway
[79]
BMF
WB
RT-PCR
LA
overexpressio
nof
miR-125bredu
cese
xpressionof
cellapop
tosis
related
proteinBc
l-2mod
ifying
factor
(Bmf)bytargetingBM
F[80]
BMPR
1BLA
miR-125btargetsB
MPR
1Bandcauses
riskin
breastcancer
pathogenesis
[81]
HuR
WB
HuR
isas
tress-in
ducedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
miR-125atargetsHuR
[82]
120581B-Ra
s2LA
miR-125bdo
wnregulates
kapp
aB-Ras2gene
expressio
n[83]
E2F3
WB
RT-PCR
LA
E2F3anindu
cero
fbladd
ercancerissup
pressedby
miR-125batproteinlevelthrou
ghregulation
ofG1S
transitionthroug
htheE
2F3-Cy
clinA2sig
nalin
gpathway
[84]
PUMA
LAmiR-125bdirectlyrepressesP
umaan
apop
tosis
regulator
inp53pathway
[79]
BAK1
CDC2
5C
PPP1CA
PPP
2CAP
RKRA
TD
GTP5
3ZA
C1LA
miR-125bdirectlyrepressesa
poptosisregulatorssuchas
BAK1
CDC2
5CP
PP1C
AP
RKRA
and
TP53AZ
AC1in
p53pathway
[79]
PPP2
CART
-PCR
LA
miR-125bdirectlyinhibitsapop
tosis
bydo
wnregulatingPP
P2CA
expressio
n[85]
TDG
qRT-PC
RmiR-125bdirectlysupp
resses
thee
xpressionof
TDGin
p53pathway
[86]
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
370 Disease Markers
miRNA gene
Pol23
RISCloading
complex
Degradation
GW182
Ago
RISCcomplex
Microprocessorcomplex
Drosha
DGCR8
DicerTRBP
Translationrepression
mRNA
Exportin 5
Inhibition of mRNA deadenylationtranslation initiation
(GW182 TRBPFMPR1 Ago Dicer
PABP eIF6)
translation elongation(GW182 TRBP
FMPR1 Ago DicerPABP)
(GW182 TRBP AgoFMPR1 Dicer
PABP CCR4 CAFPan23)
Inhibition of
AAA3 998400
5 998400
Ago
Pre-miRNA
Pri-miRNA
Pre-miRNA
Duplex miRNA
Figure 1 Biogenesis and function of microRNAs RNA polymerase 23 binds to the promoter region of specific DNA sequence and forms ahair pin structure of the pri-miRNA DGCR8 (Pasha) associates with protein Drosha which process pri-miRNA to pre-miRNA (by cleavingnucleotides from hair pin) Pre-miRNA is exported into cytoplasmwith shuttle protein exportin Inmammals the pre-miRNA is loaded ontoa multiprotein complex consisting minimally of Dicer Tar RNA Binding Protein (TRBP) and Ago2 The RNase III enzyme Dicer processesthe precursor by cleaving the stem loop to produce thematuremiRNA (canonical pathway) [11ndash13]The precursor can be loaded directly ontoAgo2 protein and this Argonaute cleaves the precursor RNA of this miRNA [13] The 51015840 nucleotide of the miRNA guide strand especially 51015840U functions as an additive anchor and helps Ago proteins to load mismatch-containing miRNAmiRNA duplexes [14] Of the two strandsthe guide strand is integrated with RISC forming miRNA-RISC complex and other strand is degraded [13ndash16] RISC is involved in inhibitionof translation initiation inhibition of translation elongation or mRNA deadenylation [15ndash17]
miRNAs in breast cancer bonemetastases and other cancerssuggesting that miRNAs could be a valuable biomarker forcancers However the regulatory mechanisms of miRNAs inbreast cancer and bone metastasis remain unclear Here wehave systematically summarized the experimentally validatedtargets of up- and downregulated miRNAs along with theirregulation in breast cancer and bonemetastasis for diagnosticand therapeutic purposes
12 MicroRNAs Synthesis MicroRNAs (miRNAs) are a classof tiny noncoding endogenous RNA molecules only 18ndash25nucleotides long These small molecules have been shownto play critical regulatory roles in a wide range of bio-logical and pathological processes miRNAs may regulatecellular gene expression at the posttranscriptional level bysuppressing translation of protein coding genes or cleavingtarget mRNAs to induce their degradation through imperfectpairing with target mRNAs of protein coding genes at 31015840UTR(untranslated region) The interacting region of 51015840 end ofmiRNA nucleotides (2 to 8 nt) is called seed sequence [18ndash20] However few reports emphasized that miRNAs can alsotarget at 51015840UTRs and protein coding regions of mRNA [1819 21 22] According to miRBase 2578 mature and 1872precursor forms of miRNAs have been identified in human(httpwwwmirbaseorg) MicroRNAs are transcribed with
RNA polymerase II (Figure 1) The RNA polymerase bindsto the promoter region of specific DNA sequence and formsa hair pin structure of the pri-miRNA The transcript thusobtained is methylated at 51015840 region and polyadenylatedat 31015840 region which undergoes posttranscriptional splicingresulting in pri-miRNA If the stem loop is found in 31015840UTRthe transcript serves as pri-miRNA [11 18] The pri-miRNA(poly or monocystronic) has about 1 kb nucleotides in lengthand its double stranded region of hair pin structure isrecognized by nuclear protein DiGeorge syndrome criticalregion 8 (DGCR8or ldquoPashardquo in invertebrates) [11 12] DGCR8associates with protein Drosha which process pri-miRNAto pre-miRNA (by cleaving nucleotides from hair pin) Pre-miRNAs (sim70 nt) have 31015840 overhang due to cleavage by RNApol II Pre-miRNA is exported into cytoplasm with shuttleprotein exportin This is an energy coupled transport withGTP bound Ran protein a nuclear transport receptor [13]
In cytoplasm the Dicer an RNase III enzyme cleaves pre-miRNA to give double strandedmiRNAduplex [14] Inmam-mals the pre-miRNA is loaded onto a multiprotein complexconsisting minimally of Dicer Tar RNA Binding Protein(TRBP) and Ago2 The Dicer processes the precursor bycleaving the stem loop to produce the mature miRNA Ago2binds with mature miRNA to form the RISC (RNA inducedsilencing complex) complex to target specific mRNA This iscanonical miRNA biogenesis (Figure 1) miRNA biogenesis
Disease Markers 371
Sample typeNormalTumor
T20
T22
T43
T17
T18
T25T4T31
T27
T35
T24
T51T5T38T2T42
T13
T29
T36
T40
T26T1T14
T37
N1
N4
T34
T49
T16
T46
T28
T21
T30T9T6T45
T48
T12
N2
T50
T15
N3
T23T8T32
T33
T47
T41
T39T3T19
T10T7T44
T11
Sample type
hsa-miR-27ahsa-miR-21hsa-miR-27bhsa-miR-31hsa-miR-205hsa-miR-206hsa-miR-125bhsa-miR-125a-5phsa-miR-10bhsa-miR-155
hsa-miR-146ahsa-miR-146b-5p
Figure 2 Heat map of differential expression of miRNAs in human normal and tumor breast cells Differential expression of miR-206 miR-31 miR-27a miR-21 miR-27b miR-205 miR-125b miR-125a-5p miR-10b miR-155 miR-146b-5p and miR-146a was identified in normalbreast and breast tumor cells by miRNA body map along with the hierarchical cluster analysis Expression of miRNAs is represented as blue(downregulated) red (upregulated) and white colors (no significant change or absence of data) Here T represents breast tumor cells and Nrepresents breast normal cells
can also depend on Ago2 protein instead of Dicer In caseof miR-451 the precursor is loaded directly onto Ago2 andthis protein cleaves the pre-miRNA The mature miR-451 isbound by Ago2 to form the RISC (RNA induced silencingcomplex) that targets erythropoietic-specific mRNAs [15]The 51015840 phosphate of the miRNA strand is essential for duplexloading into Ago and the preferred 51015840 nucleotide (uridine)of the miRNA strand (guide strand) and the base pairingstatus in the seed region and the middle of the 31015840 regionfunction as additive anchors to Ago [16] Of the two strandsthe guide strand is integratedwithRISC andother strand thatis passenger stand is degraded due to its instability [16 17]The miRNARISC complex attaches to the messenger RNA(mRNA) in one of the two ways (1) When the sequencesare perfectly complementary the miRNARISC complexbinds tightly to the mRNA and via the enzyme Ago2 themRNA is degraded [23]The poly A binding protein (PABP-)dependent poly(A) nuclease 2 (Pan2-) Pan3 deadenylases areinvolved in mRNA deadenylation that could lead to mRNAdegradation [16 18 23] Poly A seems to give stability to themRNA (2) More commonly when the sequences are imper-fectly complementary the miRNARISC complex binds andinhibits translation of the mRNA without degradation Thefinal outcome of either of these pathways is a decrease in theprotein level of the target gene The question of whether amiRNA induces translation inhibition ormRNA degradationwould depend on the level of individual miRNA specifictargets and cell backgrounds [18 19] (Figure 1)
2 miRNAs Expression
MicroRNAs play an important role in normal functioningof the cells Deregulation of miRNAs has been associated
with several disease conditions [19] Most of the miRNAsare either upregulated or downregulated thereby acting asoncogenes or tumor suppressor genes by regulating their tar-get genes To evaluate the differential expression of preferredmiRNAs between normal and breast cancer cells we per-formed miRNA body map analysis (httpwwwmirnabody-maporg) based on database instructions [24] This databaseholds expression of miRNAs from normal and diseasedtissues by RT-qPCR analysis and the data allows the analysisof differential expression of miRNAs between specific tissuesor samples The heat map provided here emphasizes thedifferential expression of miR-206 miR-31 miR-27a miR-21miR-27b miR-205 miR-125b miR-125a-5p miR-10b miR-155 miR-146b-5p and miR-146a in 51 breast cancer samplesand 4 normal breast samples of human (Figure 2) Theheat map based on color (blue down regulation and redup regulation) and cluster clearly depicts the differentialexpression of preferred miRNAs between normal and tumorbreast cellsThe up- or downregulatedmiRNAs are correlatedwith the expression of their targeted genes which couldprovide valuable physiological and pathological informationabout their systems [18ndash20]
21 Oncogenic miRNAs
211 miR-21 In breast cancer miR-21 was found to beoverexpressed [10 25] The proteomic analysis revealed thatTPM1 (Tropomysin) expression is reduced in tumor cellsand it is a target gene of miR-21 [25] TPM1 is a tumorsuppressor gene which regulates microfilament organizationand anchorage-independent growth [26] It was found thatthere is a putative miR-21 binding site at the 31015840UTR of TPM1variants When TPM1 was overexpressed in breast cancer
372 Disease Markers
(MCF-7) cells the anchorage-independent growth was sup-pressed [25] A tumor suppressor gene phosphatase andtensin homolog (PTEN) is also targeted by miR-21 [27] Up-regulation of miR-21 limits the activity of PTEN and PTENis known to limit the activity of PI3K pathway [28] Anothertumor suppressor gene product PDCD4 (programmed celldeath) was originally characterized as an inhibitor of cellulartransformation in a mouse cell culture model [29] Studieshave shown that the levels of PDCD4 mRNA as well asprotein were upregulated by miR-21 inhibition indicatingthat PDCD4 is an important functional target for miR-21[30 31] In all-transretinoic acid (ATRA) treatment miR-21 was selectively induced in ER-120572 positive cells and not inER-120572 negative cells Based on differential expression of genesregulated by ATRA in ER-120572 positive and ER-120572 negative cellsthree more direct miR-21 targets (proinflammatory cytokine(IL1B) adhesion molecule (ICAM-1 and PLAT) tissue-typeplasminogen activator (tPA)) have been predicted [32] miR-21 downregulatedMSH2 and SMAD7 in TGF-120573 pathway andcaused breast cancer progression and upregulated humanepidermal growth factor receptor 2 and thus it acted asoncogene [33 34] miR-21 with numerous gene targets inbreast cancer progression is positively regulated by a proteinnamed nucleolin (NCL) a major nucleolar protein at post-transcriptional level Overall it is emphasized that miR-21 isone of the important miRNAs involved in the regulation ofcancer cells [35 36]
212 miR-10b In metastatic breast cancer cell lines miR-10b was upregulated after activation of twist transcriptionfactor and its overexpression increased with tumor size andinvasiveness It appeared that miR-10b inhibits translationof homeobox D10 (HOXD10) resulting in induction of theprometastatic gene product RHOC [37] This RHOC inturn favors cell migration and invasion The initiation ofmetastasis in nonmetastatic cancer with overexpression ofmiR-10b in an in vivo experiment proved its role in themetastasis of breast cancer Interestingly it was identified thatmiR-10b was down-regulated in breast cancer cells comparedto normal breast cells providing evidence that miR-10b doesnot play any role during initial tumor development [38] andproliferation [39] In contrast to its oncogenic role it wasalso identified that miR-10b targeted various oncogenic genessuch as FLT1 BDNF and SHC1 a signal transduction factor[10] The targets of miR-10b identified are BUB1 PLK1 andCCNA2 [40] miR-10b also targets syndecan-1 and promotesbreast cancer metastasis and invasiveness [41]
213 miR-125 Data reported that miR-125 is likely to downregulate genes which promote tumorigenesis Overexpres-sion of miR-125a or mir-125b in an erb2-dependent cancercell line (skbr3) suppressed her2 and her3 transcript andprotein levels which decreased cell motility and invasiveness[42] miR-125a and miR-125b were significantly downregu-lated in her2-positive breast cancers Computation analysisconfirmed target sites at the 31015840UTR regions of her2 and her3for these miRNAs [43] The role of miR-125 as an oncogene
has been validated by its role in suppressing various tumorsuppressor genes including p53 gene [44]
214 miR-155 miR-155 has been shown as over-expressed inbreast cancer and further it is upregulated in normal mousemammary gland epithelial cells (NMuMG cells) by the TGF-120573Smad4 pathway and it mediates TGF-120573-induced EMT andcell invasion [10 45 46] The ectopic expression of miR-155in NMuMG cells disrupted proper tight junction formationand promoted cellmigration and invasion Conversely antag-onizing miR-155 in NMuMG cells reduced the occurrenceof TGF-120573-induced EMT and cell migration and invasionmiR-155 directly inhibits the expression of RhoA a gene thatregulates many cellular processes including cell adhesionmotility and polarity and is an important modulator ofcell junction formation and stability It is speculated thatTGF-120573Smad4 regulates expression of miR-155 resulting indown regulation of RhoA protein expression to drive EMTprogression In another study it was shown that miR-155 isassociated with cancer invasiveness in human primary breastcarcinoma that is miR-155 is highly expressed in invasivetumors but not in noninvasive cancer tissues [45 46]
Ectopic expression ofmiR-155 induces cell survival but itsknockdown leads cell to apoptosis and enhances chemosen-sitivity FOXO3a has been identified as direct target formiR-155 Inverse correlation of FOXO3a and miR-155 hasbeen observed in breast cancer cell lines and tumors miR-155 directly interacts with 31015840UTR of FOXO3a and blocksits translation Hence FOXO3a is negatively regulated bymiR-155 and FOXO3a mediates miR-155 function in thecontrol of breast cancer cell survival and growth [47]Further the phenomenon of up regulation of hexokinase2 (hk2) which enhances the metabolic activity of breastcancer cells by miR-155 makes it as a potential candidate foranticancer therapy miR-155 first activates signal transducerand activator of transcription 3 (STAT3) a transcriptionalactivator for hk2 and secondly target CEBP120573 [48] It hasbeen shown that tumor-associated circulating miRs (miR-10bmiR-34amiR-141 andmiR-155) are elevated in the bloodof breast cancer patients and hence there would be feasibilityand clinical utility of circulating miRNAs as biomarkersfor the detection and staging of breast cancer [49] It wasevident that miR-155 inhibited BMP2- BMP6- and BMP7and induced ID3 expression by targeting SMAD1 SMAD5HIVEP2 CEBPB RUNX2 and MYO10 components of theBMP signaling cascade [50] Recent findings indicate thatmiR-155 contributes to cell proliferation by down regulationof TP53INP1 in estradiol promoted breast cancer cells [51]miR-155 also downregulated tumor suppressor genes such asp53 [52] CDC73 [53] and VHL [54] in breast cancer cells InTable 1 the detailed information about oncogenic miRNAstheir target genes and their regulation during breast cancerprogression is provided
22 Tumor Suppressor miRNAs
221 miR-206 A differential expression of miR-206 wasobserved in cancer and normal tissues miR-206 targets
Disease Markers 373
Table1Targetso
foncogenicmiRNAs
andtheirroleincancer
cells
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-21
TPM1
WB
LAmiR-21d
ownregulates
TPM1inbreastcancer
andleadstoanchorage-independ
entg
rowth
[25]
E2F1
WB
RT-PCR
miR-21regulates
E2F1
byTG
F-120573sig
nalin
gpathway
[55]
TGFB
R2WB
RT-PCR
miR-21regulates
TGFB
R2by
TGF-120573sig
nalin
gpathway
[55]
TIMP3
WB
RT-PCR
TIMP3
isaM
MPinhibitormiR-21sup
pressesT
IMP3
byprom
otinginvasiv
enesso
fcancerc
ells
[56]
SERP
INB5
WB
LADow
nregulationof
PDCD
4andmaspin(SER
PINB5
)bymiR-21causestum
orogenesis
[57]
PDCD
4WB
RT-PCR
LA
miR-21d
ecreases
PDCD
4proteinam
ountsa
ndincreasesinvasion
[58]
FASCD
K6P
DCD
4CD
KN1A
FAM3C
HIPK3
PR
RG4AC
TA2BT
G2
BMPR
2SE
SN1IL6R
SOCS
5GLC
CI1APA
F1
SLC1
6A10SGK3
RP2
CFL
2
RT-PCR
M
Inhibitio
nof
miR-21inMCF
-7breastcancer
cells
show
sreduced
expressio
nof
p53tumor
supp
ressor
protein
FASCD
K6C
DKN
1AFAM3C
HIPK3
PRR
G4AC
TA2BT
G2BM
PR2
SESN
1IL6R
andtumor
supp
ressor
proteinProgrammed
CellD
eath
4(PDCD
4)area
lsodo
wnregulated
bymiR-21
[30]
FASTIMP3
WB
LADow
nregulationof
miR-21increases
FasligandandTIMP3
expressio
ntherebyitisinvolved
intheu
pregulationof
apop
tosis
[59]
CDC2
5AWB
RT-PCR
LA
miR-21sup
pressesC
dc25Aexpressio
nac
ellcycleregulator
[60]
RECK
RT-PCR
miR-21sup
pressesR
ECKandinhibitorsof
MMPs
andthus
prom
otes
invasiv
enesso
fcancerc
ells
[56]
MTA
PLA
Apop
totic
functio
nof
MTA
Pissupp
ressed
bymiR-21
[61]
RECK
WB
RT-PCR
LA
RECK
regu
latingthem
embrane-anchored
protease
isdo
wnregu
lated
bymiR-21
[62]
PDCD
4WB
LAmiR-21p
ositivelyregulates
PDCD
4throug
hinteractionwith
ovarianste
roidsa
ndTG
F-120573
signalin
gpathway
[55]
BTG2
EGFPW
BFluo
rescence
Intensity
Expressio
nof
miR-21leads
tolowe
rlevelof
BTG2which
isac
ellcycleregu
latora
ndtumor
supp
ressorItcauseslosso
fcon
trolonG1toSph
asetransition
[63]
PTEN
LAmiR-21regulates
MMP-2expressio
nin
cardiacfi
brob
lasts
oftheinfarctzone
viaP
TENpathway
[64]
BTG2
WB
RT-PCR
LA
miR-21d
irectlyinhibitsBT
G2which
isac
ellcycleinhibitorleading
toun
controlledDNA
synthesis
byactiv
ationof
forkhead
boxM1
[65]
PELI1
RT-PCR
LA
miR-21o
verexpressioninhibitsPeli1
viak
appaBsig
nalling
[66]
PDCD
4WB
Deregulationof
miR-21b
yvinylcarbam
ateind
uces
mou
selung
tumorigenesisby
downregulating
PDCD
4[67]
BIG-H
3LA
miR-21targetsbig-h3Itcou
ldbe
indu
cedby
TGF-120573anditregu
latesT
GF-120573sig
nalling
positively
andnegativ
ely
[68]
BIM
WB
LABima
apop
totic
activ
atorisc
ollectively
targeted
bymiR-21miR-24andmiR-221
andits
actio
nis
supp
ressed
[69]
HER
2WB
RT-PCR
miR-21u
pregulates
human
epidermalgrow
thfactor
receptor
2andactsas
oncogene
[34]
MSH
2SM
AD7
WB
RT-PCR
LA
miR-21d
ownregulates
MSH
2andSM
AD7in
TGF-120573pathway
andcauses
breastcancer
progression
[33]
374 Disease Markers
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-10b
HOXD
10WB
LAOverexpressionof
miR-10b
indu
cesrob
ustinvasionandmetastasis
bydo
wnregu
latin
gtransla
tionof
homeobo
xD10
[37]
KLF4
WB
RT-PCR
LA
miR-10b
redu
cese
ndogenou
sKLF
4protein
atum
orsupp
ressor
[70]
SFRS
1WB
RT-PCR
LA
miR-10b
supp
resses
SFRS
1(SR
-family
splicingfactor)a
ndcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
SFRS
10LA
miR-10b
supp
resses
SFRS
10andcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
miR-125a-5p
ERBB
2WB
RT-PCR
LA
Expressio
nof
either
miR-125ao
rmiR-125bresults
insupp
ressionof
ERBB
2andER
BB3
(oncogenefam
ily)a
ndexhibitsredu
cedmigratio
nandinvasio
ncapacitie
s[72]
TP53
WB
RT-PCR
LA
miR-125binhibitsp53gene
transla
tionally
andtranscrip
tionally
andtheisomer
ofmiRNA-
125
(miRNA-
125a)translationally
arrests
mRN
Aof
thep
53a
tumor
supp
ressor
gene
[44]
HuR
WB
RT-PCR
LA
HuR
isas
tressindu
cedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
Expressio
nof
miR-125as
ignificantly
redu
cesH
uR
[73]
ARID3B
WB
ARID3B
isoverexpressedin
human
ovariancancerO
verexpressionof
miR-125aind
uces
conversio
nof
high
lyinvasiv
eovaria
ncancer
cells
from
amesenchym
alto
anepith
elial
morph
ologyandsupp
resses
cancer
bynegativ
elyregu
latin
gEM
T[74]
miR-125b
BAK1
WB
RT-PCR
LAM
Overexpressionof
miR-125bstimulates
androgen-in
depend
entg
rowth
ofprostatecancer
bydo
wnregu
latin
gBa
k1(proapop
totic
regu
lator)
[75]
CDK6
CDC2
5AWB
miR-125boverexpressio
ndecreasesC
DK6
andCD
C25A
expressio
nItregu
latesp
roliferationof
U251g
liomas
tem
cells
throug
hcellcycle
regu
latedproteins
CDK6
andCD
C25A
[76]
SMO
WB
RT-PCR
LA
Dow
nregulationof
miR-125ballowsh
ighlevelsof
Hedgeho
g-depend
entgenee
xpressionleading
totumor
cellproliferatio
nby
allowingHedgeho
gsig
nalling
[77]
ST18
WB
BranchedL
AD
NA
Prob
eAssay
ST18
isdo
wnregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
DICER
1WB
LAB
ranchedDNA
Prob
eAssay
DICER
1isd
ownregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
TP53IN
P1LA
miR-125bdirectlyrepressesT
P53INP1anapop
tosis
regulator
inp53pathway
[79]
BMF
WB
RT-PCR
LA
overexpressio
nof
miR-125bredu
cese
xpressionof
cellapop
tosis
related
proteinBc
l-2mod
ifying
factor
(Bmf)bytargetingBM
F[80]
BMPR
1BLA
miR-125btargetsB
MPR
1Bandcauses
riskin
breastcancer
pathogenesis
[81]
HuR
WB
HuR
isas
tress-in
ducedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
miR-125atargetsHuR
[82]
120581B-Ra
s2LA
miR-125bdo
wnregulates
kapp
aB-Ras2gene
expressio
n[83]
E2F3
WB
RT-PCR
LA
E2F3anindu
cero
fbladd
ercancerissup
pressedby
miR-125batproteinlevelthrou
ghregulation
ofG1S
transitionthroug
htheE
2F3-Cy
clinA2sig
nalin
gpathway
[84]
PUMA
LAmiR-125bdirectlyrepressesP
umaan
apop
tosis
regulator
inp53pathway
[79]
BAK1
CDC2
5C
PPP1CA
PPP
2CAP
RKRA
TD
GTP5
3ZA
C1LA
miR-125bdirectlyrepressesa
poptosisregulatorssuchas
BAK1
CDC2
5CP
PP1C
AP
RKRA
and
TP53AZ
AC1in
p53pathway
[79]
PPP2
CART
-PCR
LA
miR-125bdirectlyinhibitsapop
tosis
bydo
wnregulatingPP
P2CA
expressio
n[85]
TDG
qRT-PC
RmiR-125bdirectlysupp
resses
thee
xpressionof
TDGin
p53pathway
[86]
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 371
Sample typeNormalTumor
T20
T22
T43
T17
T18
T25T4T31
T27
T35
T24
T51T5T38T2T42
T13
T29
T36
T40
T26T1T14
T37
N1
N4
T34
T49
T16
T46
T28
T21
T30T9T6T45
T48
T12
N2
T50
T15
N3
T23T8T32
T33
T47
T41
T39T3T19
T10T7T44
T11
Sample type
hsa-miR-27ahsa-miR-21hsa-miR-27bhsa-miR-31hsa-miR-205hsa-miR-206hsa-miR-125bhsa-miR-125a-5phsa-miR-10bhsa-miR-155
hsa-miR-146ahsa-miR-146b-5p
Figure 2 Heat map of differential expression of miRNAs in human normal and tumor breast cells Differential expression of miR-206 miR-31 miR-27a miR-21 miR-27b miR-205 miR-125b miR-125a-5p miR-10b miR-155 miR-146b-5p and miR-146a was identified in normalbreast and breast tumor cells by miRNA body map along with the hierarchical cluster analysis Expression of miRNAs is represented as blue(downregulated) red (upregulated) and white colors (no significant change or absence of data) Here T represents breast tumor cells and Nrepresents breast normal cells
can also depend on Ago2 protein instead of Dicer In caseof miR-451 the precursor is loaded directly onto Ago2 andthis protein cleaves the pre-miRNA The mature miR-451 isbound by Ago2 to form the RISC (RNA induced silencingcomplex) that targets erythropoietic-specific mRNAs [15]The 51015840 phosphate of the miRNA strand is essential for duplexloading into Ago and the preferred 51015840 nucleotide (uridine)of the miRNA strand (guide strand) and the base pairingstatus in the seed region and the middle of the 31015840 regionfunction as additive anchors to Ago [16] Of the two strandsthe guide strand is integratedwithRISC andother strand thatis passenger stand is degraded due to its instability [16 17]The miRNARISC complex attaches to the messenger RNA(mRNA) in one of the two ways (1) When the sequencesare perfectly complementary the miRNARISC complexbinds tightly to the mRNA and via the enzyme Ago2 themRNA is degraded [23]The poly A binding protein (PABP-)dependent poly(A) nuclease 2 (Pan2-) Pan3 deadenylases areinvolved in mRNA deadenylation that could lead to mRNAdegradation [16 18 23] Poly A seems to give stability to themRNA (2) More commonly when the sequences are imper-fectly complementary the miRNARISC complex binds andinhibits translation of the mRNA without degradation Thefinal outcome of either of these pathways is a decrease in theprotein level of the target gene The question of whether amiRNA induces translation inhibition ormRNA degradationwould depend on the level of individual miRNA specifictargets and cell backgrounds [18 19] (Figure 1)
2 miRNAs Expression
MicroRNAs play an important role in normal functioningof the cells Deregulation of miRNAs has been associated
with several disease conditions [19] Most of the miRNAsare either upregulated or downregulated thereby acting asoncogenes or tumor suppressor genes by regulating their tar-get genes To evaluate the differential expression of preferredmiRNAs between normal and breast cancer cells we per-formed miRNA body map analysis (httpwwwmirnabody-maporg) based on database instructions [24] This databaseholds expression of miRNAs from normal and diseasedtissues by RT-qPCR analysis and the data allows the analysisof differential expression of miRNAs between specific tissuesor samples The heat map provided here emphasizes thedifferential expression of miR-206 miR-31 miR-27a miR-21miR-27b miR-205 miR-125b miR-125a-5p miR-10b miR-155 miR-146b-5p and miR-146a in 51 breast cancer samplesand 4 normal breast samples of human (Figure 2) Theheat map based on color (blue down regulation and redup regulation) and cluster clearly depicts the differentialexpression of preferred miRNAs between normal and tumorbreast cellsThe up- or downregulatedmiRNAs are correlatedwith the expression of their targeted genes which couldprovide valuable physiological and pathological informationabout their systems [18ndash20]
21 Oncogenic miRNAs
211 miR-21 In breast cancer miR-21 was found to beoverexpressed [10 25] The proteomic analysis revealed thatTPM1 (Tropomysin) expression is reduced in tumor cellsand it is a target gene of miR-21 [25] TPM1 is a tumorsuppressor gene which regulates microfilament organizationand anchorage-independent growth [26] It was found thatthere is a putative miR-21 binding site at the 31015840UTR of TPM1variants When TPM1 was overexpressed in breast cancer
372 Disease Markers
(MCF-7) cells the anchorage-independent growth was sup-pressed [25] A tumor suppressor gene phosphatase andtensin homolog (PTEN) is also targeted by miR-21 [27] Up-regulation of miR-21 limits the activity of PTEN and PTENis known to limit the activity of PI3K pathway [28] Anothertumor suppressor gene product PDCD4 (programmed celldeath) was originally characterized as an inhibitor of cellulartransformation in a mouse cell culture model [29] Studieshave shown that the levels of PDCD4 mRNA as well asprotein were upregulated by miR-21 inhibition indicatingthat PDCD4 is an important functional target for miR-21[30 31] In all-transretinoic acid (ATRA) treatment miR-21 was selectively induced in ER-120572 positive cells and not inER-120572 negative cells Based on differential expression of genesregulated by ATRA in ER-120572 positive and ER-120572 negative cellsthree more direct miR-21 targets (proinflammatory cytokine(IL1B) adhesion molecule (ICAM-1 and PLAT) tissue-typeplasminogen activator (tPA)) have been predicted [32] miR-21 downregulatedMSH2 and SMAD7 in TGF-120573 pathway andcaused breast cancer progression and upregulated humanepidermal growth factor receptor 2 and thus it acted asoncogene [33 34] miR-21 with numerous gene targets inbreast cancer progression is positively regulated by a proteinnamed nucleolin (NCL) a major nucleolar protein at post-transcriptional level Overall it is emphasized that miR-21 isone of the important miRNAs involved in the regulation ofcancer cells [35 36]
212 miR-10b In metastatic breast cancer cell lines miR-10b was upregulated after activation of twist transcriptionfactor and its overexpression increased with tumor size andinvasiveness It appeared that miR-10b inhibits translationof homeobox D10 (HOXD10) resulting in induction of theprometastatic gene product RHOC [37] This RHOC inturn favors cell migration and invasion The initiation ofmetastasis in nonmetastatic cancer with overexpression ofmiR-10b in an in vivo experiment proved its role in themetastasis of breast cancer Interestingly it was identified thatmiR-10b was down-regulated in breast cancer cells comparedto normal breast cells providing evidence that miR-10b doesnot play any role during initial tumor development [38] andproliferation [39] In contrast to its oncogenic role it wasalso identified that miR-10b targeted various oncogenic genessuch as FLT1 BDNF and SHC1 a signal transduction factor[10] The targets of miR-10b identified are BUB1 PLK1 andCCNA2 [40] miR-10b also targets syndecan-1 and promotesbreast cancer metastasis and invasiveness [41]
213 miR-125 Data reported that miR-125 is likely to downregulate genes which promote tumorigenesis Overexpres-sion of miR-125a or mir-125b in an erb2-dependent cancercell line (skbr3) suppressed her2 and her3 transcript andprotein levels which decreased cell motility and invasiveness[42] miR-125a and miR-125b were significantly downregu-lated in her2-positive breast cancers Computation analysisconfirmed target sites at the 31015840UTR regions of her2 and her3for these miRNAs [43] The role of miR-125 as an oncogene
has been validated by its role in suppressing various tumorsuppressor genes including p53 gene [44]
214 miR-155 miR-155 has been shown as over-expressed inbreast cancer and further it is upregulated in normal mousemammary gland epithelial cells (NMuMG cells) by the TGF-120573Smad4 pathway and it mediates TGF-120573-induced EMT andcell invasion [10 45 46] The ectopic expression of miR-155in NMuMG cells disrupted proper tight junction formationand promoted cellmigration and invasion Conversely antag-onizing miR-155 in NMuMG cells reduced the occurrenceof TGF-120573-induced EMT and cell migration and invasionmiR-155 directly inhibits the expression of RhoA a gene thatregulates many cellular processes including cell adhesionmotility and polarity and is an important modulator ofcell junction formation and stability It is speculated thatTGF-120573Smad4 regulates expression of miR-155 resulting indown regulation of RhoA protein expression to drive EMTprogression In another study it was shown that miR-155 isassociated with cancer invasiveness in human primary breastcarcinoma that is miR-155 is highly expressed in invasivetumors but not in noninvasive cancer tissues [45 46]
Ectopic expression ofmiR-155 induces cell survival but itsknockdown leads cell to apoptosis and enhances chemosen-sitivity FOXO3a has been identified as direct target formiR-155 Inverse correlation of FOXO3a and miR-155 hasbeen observed in breast cancer cell lines and tumors miR-155 directly interacts with 31015840UTR of FOXO3a and blocksits translation Hence FOXO3a is negatively regulated bymiR-155 and FOXO3a mediates miR-155 function in thecontrol of breast cancer cell survival and growth [47]Further the phenomenon of up regulation of hexokinase2 (hk2) which enhances the metabolic activity of breastcancer cells by miR-155 makes it as a potential candidate foranticancer therapy miR-155 first activates signal transducerand activator of transcription 3 (STAT3) a transcriptionalactivator for hk2 and secondly target CEBP120573 [48] It hasbeen shown that tumor-associated circulating miRs (miR-10bmiR-34amiR-141 andmiR-155) are elevated in the bloodof breast cancer patients and hence there would be feasibilityand clinical utility of circulating miRNAs as biomarkersfor the detection and staging of breast cancer [49] It wasevident that miR-155 inhibited BMP2- BMP6- and BMP7and induced ID3 expression by targeting SMAD1 SMAD5HIVEP2 CEBPB RUNX2 and MYO10 components of theBMP signaling cascade [50] Recent findings indicate thatmiR-155 contributes to cell proliferation by down regulationof TP53INP1 in estradiol promoted breast cancer cells [51]miR-155 also downregulated tumor suppressor genes such asp53 [52] CDC73 [53] and VHL [54] in breast cancer cells InTable 1 the detailed information about oncogenic miRNAstheir target genes and their regulation during breast cancerprogression is provided
22 Tumor Suppressor miRNAs
221 miR-206 A differential expression of miR-206 wasobserved in cancer and normal tissues miR-206 targets
Disease Markers 373
Table1Targetso
foncogenicmiRNAs
andtheirroleincancer
cells
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-21
TPM1
WB
LAmiR-21d
ownregulates
TPM1inbreastcancer
andleadstoanchorage-independ
entg
rowth
[25]
E2F1
WB
RT-PCR
miR-21regulates
E2F1
byTG
F-120573sig
nalin
gpathway
[55]
TGFB
R2WB
RT-PCR
miR-21regulates
TGFB
R2by
TGF-120573sig
nalin
gpathway
[55]
TIMP3
WB
RT-PCR
TIMP3
isaM
MPinhibitormiR-21sup
pressesT
IMP3
byprom
otinginvasiv
enesso
fcancerc
ells
[56]
SERP
INB5
WB
LADow
nregulationof
PDCD
4andmaspin(SER
PINB5
)bymiR-21causestum
orogenesis
[57]
PDCD
4WB
RT-PCR
LA
miR-21d
ecreases
PDCD
4proteinam
ountsa
ndincreasesinvasion
[58]
FASCD
K6P
DCD
4CD
KN1A
FAM3C
HIPK3
PR
RG4AC
TA2BT
G2
BMPR
2SE
SN1IL6R
SOCS
5GLC
CI1APA
F1
SLC1
6A10SGK3
RP2
CFL
2
RT-PCR
M
Inhibitio
nof
miR-21inMCF
-7breastcancer
cells
show
sreduced
expressio
nof
p53tumor
supp
ressor
protein
FASCD
K6C
DKN
1AFAM3C
HIPK3
PRR
G4AC
TA2BT
G2BM
PR2
SESN
1IL6R
andtumor
supp
ressor
proteinProgrammed
CellD
eath
4(PDCD
4)area
lsodo
wnregulated
bymiR-21
[30]
FASTIMP3
WB
LADow
nregulationof
miR-21increases
FasligandandTIMP3
expressio
ntherebyitisinvolved
intheu
pregulationof
apop
tosis
[59]
CDC2
5AWB
RT-PCR
LA
miR-21sup
pressesC
dc25Aexpressio
nac
ellcycleregulator
[60]
RECK
RT-PCR
miR-21sup
pressesR
ECKandinhibitorsof
MMPs
andthus
prom
otes
invasiv
enesso
fcancerc
ells
[56]
MTA
PLA
Apop
totic
functio
nof
MTA
Pissupp
ressed
bymiR-21
[61]
RECK
WB
RT-PCR
LA
RECK
regu
latingthem
embrane-anchored
protease
isdo
wnregu
lated
bymiR-21
[62]
PDCD
4WB
LAmiR-21p
ositivelyregulates
PDCD
4throug
hinteractionwith
ovarianste
roidsa
ndTG
F-120573
signalin
gpathway
[55]
BTG2
EGFPW
BFluo
rescence
Intensity
Expressio
nof
miR-21leads
tolowe
rlevelof
BTG2which
isac
ellcycleregu
latora
ndtumor
supp
ressorItcauseslosso
fcon
trolonG1toSph
asetransition
[63]
PTEN
LAmiR-21regulates
MMP-2expressio
nin
cardiacfi
brob
lasts
oftheinfarctzone
viaP
TENpathway
[64]
BTG2
WB
RT-PCR
LA
miR-21d
irectlyinhibitsBT
G2which
isac
ellcycleinhibitorleading
toun
controlledDNA
synthesis
byactiv
ationof
forkhead
boxM1
[65]
PELI1
RT-PCR
LA
miR-21o
verexpressioninhibitsPeli1
viak
appaBsig
nalling
[66]
PDCD
4WB
Deregulationof
miR-21b
yvinylcarbam
ateind
uces
mou
selung
tumorigenesisby
downregulating
PDCD
4[67]
BIG-H
3LA
miR-21targetsbig-h3Itcou
ldbe
indu
cedby
TGF-120573anditregu
latesT
GF-120573sig
nalling
positively
andnegativ
ely
[68]
BIM
WB
LABima
apop
totic
activ
atorisc
ollectively
targeted
bymiR-21miR-24andmiR-221
andits
actio
nis
supp
ressed
[69]
HER
2WB
RT-PCR
miR-21u
pregulates
human
epidermalgrow
thfactor
receptor
2andactsas
oncogene
[34]
MSH
2SM
AD7
WB
RT-PCR
LA
miR-21d
ownregulates
MSH
2andSM
AD7in
TGF-120573pathway
andcauses
breastcancer
progression
[33]
374 Disease Markers
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-10b
HOXD
10WB
LAOverexpressionof
miR-10b
indu
cesrob
ustinvasionandmetastasis
bydo
wnregu
latin
gtransla
tionof
homeobo
xD10
[37]
KLF4
WB
RT-PCR
LA
miR-10b
redu
cese
ndogenou
sKLF
4protein
atum
orsupp
ressor
[70]
SFRS
1WB
RT-PCR
LA
miR-10b
supp
resses
SFRS
1(SR
-family
splicingfactor)a
ndcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
SFRS
10LA
miR-10b
supp
resses
SFRS
10andcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
miR-125a-5p
ERBB
2WB
RT-PCR
LA
Expressio
nof
either
miR-125ao
rmiR-125bresults
insupp
ressionof
ERBB
2andER
BB3
(oncogenefam
ily)a
ndexhibitsredu
cedmigratio
nandinvasio
ncapacitie
s[72]
TP53
WB
RT-PCR
LA
miR-125binhibitsp53gene
transla
tionally
andtranscrip
tionally
andtheisomer
ofmiRNA-
125
(miRNA-
125a)translationally
arrests
mRN
Aof
thep
53a
tumor
supp
ressor
gene
[44]
HuR
WB
RT-PCR
LA
HuR
isas
tressindu
cedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
Expressio
nof
miR-125as
ignificantly
redu
cesH
uR
[73]
ARID3B
WB
ARID3B
isoverexpressedin
human
ovariancancerO
verexpressionof
miR-125aind
uces
conversio
nof
high
lyinvasiv
eovaria
ncancer
cells
from
amesenchym
alto
anepith
elial
morph
ologyandsupp
resses
cancer
bynegativ
elyregu
latin
gEM
T[74]
miR-125b
BAK1
WB
RT-PCR
LAM
Overexpressionof
miR-125bstimulates
androgen-in
depend
entg
rowth
ofprostatecancer
bydo
wnregu
latin
gBa
k1(proapop
totic
regu
lator)
[75]
CDK6
CDC2
5AWB
miR-125boverexpressio
ndecreasesC
DK6
andCD
C25A
expressio
nItregu
latesp
roliferationof
U251g
liomas
tem
cells
throug
hcellcycle
regu
latedproteins
CDK6
andCD
C25A
[76]
SMO
WB
RT-PCR
LA
Dow
nregulationof
miR-125ballowsh
ighlevelsof
Hedgeho
g-depend
entgenee
xpressionleading
totumor
cellproliferatio
nby
allowingHedgeho
gsig
nalling
[77]
ST18
WB
BranchedL
AD
NA
Prob
eAssay
ST18
isdo
wnregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
DICER
1WB
LAB
ranchedDNA
Prob
eAssay
DICER
1isd
ownregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
TP53IN
P1LA
miR-125bdirectlyrepressesT
P53INP1anapop
tosis
regulator
inp53pathway
[79]
BMF
WB
RT-PCR
LA
overexpressio
nof
miR-125bredu
cese
xpressionof
cellapop
tosis
related
proteinBc
l-2mod
ifying
factor
(Bmf)bytargetingBM
F[80]
BMPR
1BLA
miR-125btargetsB
MPR
1Bandcauses
riskin
breastcancer
pathogenesis
[81]
HuR
WB
HuR
isas
tress-in
ducedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
miR-125atargetsHuR
[82]
120581B-Ra
s2LA
miR-125bdo
wnregulates
kapp
aB-Ras2gene
expressio
n[83]
E2F3
WB
RT-PCR
LA
E2F3anindu
cero
fbladd
ercancerissup
pressedby
miR-125batproteinlevelthrou
ghregulation
ofG1S
transitionthroug
htheE
2F3-Cy
clinA2sig
nalin
gpathway
[84]
PUMA
LAmiR-125bdirectlyrepressesP
umaan
apop
tosis
regulator
inp53pathway
[79]
BAK1
CDC2
5C
PPP1CA
PPP
2CAP
RKRA
TD
GTP5
3ZA
C1LA
miR-125bdirectlyrepressesa
poptosisregulatorssuchas
BAK1
CDC2
5CP
PP1C
AP
RKRA
and
TP53AZ
AC1in
p53pathway
[79]
PPP2
CART
-PCR
LA
miR-125bdirectlyinhibitsapop
tosis
bydo
wnregulatingPP
P2CA
expressio
n[85]
TDG
qRT-PC
RmiR-125bdirectlysupp
resses
thee
xpressionof
TDGin
p53pathway
[86]
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
372 Disease Markers
(MCF-7) cells the anchorage-independent growth was sup-pressed [25] A tumor suppressor gene phosphatase andtensin homolog (PTEN) is also targeted by miR-21 [27] Up-regulation of miR-21 limits the activity of PTEN and PTENis known to limit the activity of PI3K pathway [28] Anothertumor suppressor gene product PDCD4 (programmed celldeath) was originally characterized as an inhibitor of cellulartransformation in a mouse cell culture model [29] Studieshave shown that the levels of PDCD4 mRNA as well asprotein were upregulated by miR-21 inhibition indicatingthat PDCD4 is an important functional target for miR-21[30 31] In all-transretinoic acid (ATRA) treatment miR-21 was selectively induced in ER-120572 positive cells and not inER-120572 negative cells Based on differential expression of genesregulated by ATRA in ER-120572 positive and ER-120572 negative cellsthree more direct miR-21 targets (proinflammatory cytokine(IL1B) adhesion molecule (ICAM-1 and PLAT) tissue-typeplasminogen activator (tPA)) have been predicted [32] miR-21 downregulatedMSH2 and SMAD7 in TGF-120573 pathway andcaused breast cancer progression and upregulated humanepidermal growth factor receptor 2 and thus it acted asoncogene [33 34] miR-21 with numerous gene targets inbreast cancer progression is positively regulated by a proteinnamed nucleolin (NCL) a major nucleolar protein at post-transcriptional level Overall it is emphasized that miR-21 isone of the important miRNAs involved in the regulation ofcancer cells [35 36]
212 miR-10b In metastatic breast cancer cell lines miR-10b was upregulated after activation of twist transcriptionfactor and its overexpression increased with tumor size andinvasiveness It appeared that miR-10b inhibits translationof homeobox D10 (HOXD10) resulting in induction of theprometastatic gene product RHOC [37] This RHOC inturn favors cell migration and invasion The initiation ofmetastasis in nonmetastatic cancer with overexpression ofmiR-10b in an in vivo experiment proved its role in themetastasis of breast cancer Interestingly it was identified thatmiR-10b was down-regulated in breast cancer cells comparedto normal breast cells providing evidence that miR-10b doesnot play any role during initial tumor development [38] andproliferation [39] In contrast to its oncogenic role it wasalso identified that miR-10b targeted various oncogenic genessuch as FLT1 BDNF and SHC1 a signal transduction factor[10] The targets of miR-10b identified are BUB1 PLK1 andCCNA2 [40] miR-10b also targets syndecan-1 and promotesbreast cancer metastasis and invasiveness [41]
213 miR-125 Data reported that miR-125 is likely to downregulate genes which promote tumorigenesis Overexpres-sion of miR-125a or mir-125b in an erb2-dependent cancercell line (skbr3) suppressed her2 and her3 transcript andprotein levels which decreased cell motility and invasiveness[42] miR-125a and miR-125b were significantly downregu-lated in her2-positive breast cancers Computation analysisconfirmed target sites at the 31015840UTR regions of her2 and her3for these miRNAs [43] The role of miR-125 as an oncogene
has been validated by its role in suppressing various tumorsuppressor genes including p53 gene [44]
214 miR-155 miR-155 has been shown as over-expressed inbreast cancer and further it is upregulated in normal mousemammary gland epithelial cells (NMuMG cells) by the TGF-120573Smad4 pathway and it mediates TGF-120573-induced EMT andcell invasion [10 45 46] The ectopic expression of miR-155in NMuMG cells disrupted proper tight junction formationand promoted cellmigration and invasion Conversely antag-onizing miR-155 in NMuMG cells reduced the occurrenceof TGF-120573-induced EMT and cell migration and invasionmiR-155 directly inhibits the expression of RhoA a gene thatregulates many cellular processes including cell adhesionmotility and polarity and is an important modulator ofcell junction formation and stability It is speculated thatTGF-120573Smad4 regulates expression of miR-155 resulting indown regulation of RhoA protein expression to drive EMTprogression In another study it was shown that miR-155 isassociated with cancer invasiveness in human primary breastcarcinoma that is miR-155 is highly expressed in invasivetumors but not in noninvasive cancer tissues [45 46]
Ectopic expression ofmiR-155 induces cell survival but itsknockdown leads cell to apoptosis and enhances chemosen-sitivity FOXO3a has been identified as direct target formiR-155 Inverse correlation of FOXO3a and miR-155 hasbeen observed in breast cancer cell lines and tumors miR-155 directly interacts with 31015840UTR of FOXO3a and blocksits translation Hence FOXO3a is negatively regulated bymiR-155 and FOXO3a mediates miR-155 function in thecontrol of breast cancer cell survival and growth [47]Further the phenomenon of up regulation of hexokinase2 (hk2) which enhances the metabolic activity of breastcancer cells by miR-155 makes it as a potential candidate foranticancer therapy miR-155 first activates signal transducerand activator of transcription 3 (STAT3) a transcriptionalactivator for hk2 and secondly target CEBP120573 [48] It hasbeen shown that tumor-associated circulating miRs (miR-10bmiR-34amiR-141 andmiR-155) are elevated in the bloodof breast cancer patients and hence there would be feasibilityand clinical utility of circulating miRNAs as biomarkersfor the detection and staging of breast cancer [49] It wasevident that miR-155 inhibited BMP2- BMP6- and BMP7and induced ID3 expression by targeting SMAD1 SMAD5HIVEP2 CEBPB RUNX2 and MYO10 components of theBMP signaling cascade [50] Recent findings indicate thatmiR-155 contributes to cell proliferation by down regulationof TP53INP1 in estradiol promoted breast cancer cells [51]miR-155 also downregulated tumor suppressor genes such asp53 [52] CDC73 [53] and VHL [54] in breast cancer cells InTable 1 the detailed information about oncogenic miRNAstheir target genes and their regulation during breast cancerprogression is provided
22 Tumor Suppressor miRNAs
221 miR-206 A differential expression of miR-206 wasobserved in cancer and normal tissues miR-206 targets
Disease Markers 373
Table1Targetso
foncogenicmiRNAs
andtheirroleincancer
cells
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-21
TPM1
WB
LAmiR-21d
ownregulates
TPM1inbreastcancer
andleadstoanchorage-independ
entg
rowth
[25]
E2F1
WB
RT-PCR
miR-21regulates
E2F1
byTG
F-120573sig
nalin
gpathway
[55]
TGFB
R2WB
RT-PCR
miR-21regulates
TGFB
R2by
TGF-120573sig
nalin
gpathway
[55]
TIMP3
WB
RT-PCR
TIMP3
isaM
MPinhibitormiR-21sup
pressesT
IMP3
byprom
otinginvasiv
enesso
fcancerc
ells
[56]
SERP
INB5
WB
LADow
nregulationof
PDCD
4andmaspin(SER
PINB5
)bymiR-21causestum
orogenesis
[57]
PDCD
4WB
RT-PCR
LA
miR-21d
ecreases
PDCD
4proteinam
ountsa
ndincreasesinvasion
[58]
FASCD
K6P
DCD
4CD
KN1A
FAM3C
HIPK3
PR
RG4AC
TA2BT
G2
BMPR
2SE
SN1IL6R
SOCS
5GLC
CI1APA
F1
SLC1
6A10SGK3
RP2
CFL
2
RT-PCR
M
Inhibitio
nof
miR-21inMCF
-7breastcancer
cells
show
sreduced
expressio
nof
p53tumor
supp
ressor
protein
FASCD
K6C
DKN
1AFAM3C
HIPK3
PRR
G4AC
TA2BT
G2BM
PR2
SESN
1IL6R
andtumor
supp
ressor
proteinProgrammed
CellD
eath
4(PDCD
4)area
lsodo
wnregulated
bymiR-21
[30]
FASTIMP3
WB
LADow
nregulationof
miR-21increases
FasligandandTIMP3
expressio
ntherebyitisinvolved
intheu
pregulationof
apop
tosis
[59]
CDC2
5AWB
RT-PCR
LA
miR-21sup
pressesC
dc25Aexpressio
nac
ellcycleregulator
[60]
RECK
RT-PCR
miR-21sup
pressesR
ECKandinhibitorsof
MMPs
andthus
prom
otes
invasiv
enesso
fcancerc
ells
[56]
MTA
PLA
Apop
totic
functio
nof
MTA
Pissupp
ressed
bymiR-21
[61]
RECK
WB
RT-PCR
LA
RECK
regu
latingthem
embrane-anchored
protease
isdo
wnregu
lated
bymiR-21
[62]
PDCD
4WB
LAmiR-21p
ositivelyregulates
PDCD
4throug
hinteractionwith
ovarianste
roidsa
ndTG
F-120573
signalin
gpathway
[55]
BTG2
EGFPW
BFluo
rescence
Intensity
Expressio
nof
miR-21leads
tolowe
rlevelof
BTG2which
isac
ellcycleregu
latora
ndtumor
supp
ressorItcauseslosso
fcon
trolonG1toSph
asetransition
[63]
PTEN
LAmiR-21regulates
MMP-2expressio
nin
cardiacfi
brob
lasts
oftheinfarctzone
viaP
TENpathway
[64]
BTG2
WB
RT-PCR
LA
miR-21d
irectlyinhibitsBT
G2which
isac
ellcycleinhibitorleading
toun
controlledDNA
synthesis
byactiv
ationof
forkhead
boxM1
[65]
PELI1
RT-PCR
LA
miR-21o
verexpressioninhibitsPeli1
viak
appaBsig
nalling
[66]
PDCD
4WB
Deregulationof
miR-21b
yvinylcarbam
ateind
uces
mou
selung
tumorigenesisby
downregulating
PDCD
4[67]
BIG-H
3LA
miR-21targetsbig-h3Itcou
ldbe
indu
cedby
TGF-120573anditregu
latesT
GF-120573sig
nalling
positively
andnegativ
ely
[68]
BIM
WB
LABima
apop
totic
activ
atorisc
ollectively
targeted
bymiR-21miR-24andmiR-221
andits
actio
nis
supp
ressed
[69]
HER
2WB
RT-PCR
miR-21u
pregulates
human
epidermalgrow
thfactor
receptor
2andactsas
oncogene
[34]
MSH
2SM
AD7
WB
RT-PCR
LA
miR-21d
ownregulates
MSH
2andSM
AD7in
TGF-120573pathway
andcauses
breastcancer
progression
[33]
374 Disease Markers
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-10b
HOXD
10WB
LAOverexpressionof
miR-10b
indu
cesrob
ustinvasionandmetastasis
bydo
wnregu
latin
gtransla
tionof
homeobo
xD10
[37]
KLF4
WB
RT-PCR
LA
miR-10b
redu
cese
ndogenou
sKLF
4protein
atum
orsupp
ressor
[70]
SFRS
1WB
RT-PCR
LA
miR-10b
supp
resses
SFRS
1(SR
-family
splicingfactor)a
ndcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
SFRS
10LA
miR-10b
supp
resses
SFRS
10andcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
miR-125a-5p
ERBB
2WB
RT-PCR
LA
Expressio
nof
either
miR-125ao
rmiR-125bresults
insupp
ressionof
ERBB
2andER
BB3
(oncogenefam
ily)a
ndexhibitsredu
cedmigratio
nandinvasio
ncapacitie
s[72]
TP53
WB
RT-PCR
LA
miR-125binhibitsp53gene
transla
tionally
andtranscrip
tionally
andtheisomer
ofmiRNA-
125
(miRNA-
125a)translationally
arrests
mRN
Aof
thep
53a
tumor
supp
ressor
gene
[44]
HuR
WB
RT-PCR
LA
HuR
isas
tressindu
cedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
Expressio
nof
miR-125as
ignificantly
redu
cesH
uR
[73]
ARID3B
WB
ARID3B
isoverexpressedin
human
ovariancancerO
verexpressionof
miR-125aind
uces
conversio
nof
high
lyinvasiv
eovaria
ncancer
cells
from
amesenchym
alto
anepith
elial
morph
ologyandsupp
resses
cancer
bynegativ
elyregu
latin
gEM
T[74]
miR-125b
BAK1
WB
RT-PCR
LAM
Overexpressionof
miR-125bstimulates
androgen-in
depend
entg
rowth
ofprostatecancer
bydo
wnregu
latin
gBa
k1(proapop
totic
regu
lator)
[75]
CDK6
CDC2
5AWB
miR-125boverexpressio
ndecreasesC
DK6
andCD
C25A
expressio
nItregu
latesp
roliferationof
U251g
liomas
tem
cells
throug
hcellcycle
regu
latedproteins
CDK6
andCD
C25A
[76]
SMO
WB
RT-PCR
LA
Dow
nregulationof
miR-125ballowsh
ighlevelsof
Hedgeho
g-depend
entgenee
xpressionleading
totumor
cellproliferatio
nby
allowingHedgeho
gsig
nalling
[77]
ST18
WB
BranchedL
AD
NA
Prob
eAssay
ST18
isdo
wnregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
DICER
1WB
LAB
ranchedDNA
Prob
eAssay
DICER
1isd
ownregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
TP53IN
P1LA
miR-125bdirectlyrepressesT
P53INP1anapop
tosis
regulator
inp53pathway
[79]
BMF
WB
RT-PCR
LA
overexpressio
nof
miR-125bredu
cese
xpressionof
cellapop
tosis
related
proteinBc
l-2mod
ifying
factor
(Bmf)bytargetingBM
F[80]
BMPR
1BLA
miR-125btargetsB
MPR
1Bandcauses
riskin
breastcancer
pathogenesis
[81]
HuR
WB
HuR
isas
tress-in
ducedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
miR-125atargetsHuR
[82]
120581B-Ra
s2LA
miR-125bdo
wnregulates
kapp
aB-Ras2gene
expressio
n[83]
E2F3
WB
RT-PCR
LA
E2F3anindu
cero
fbladd
ercancerissup
pressedby
miR-125batproteinlevelthrou
ghregulation
ofG1S
transitionthroug
htheE
2F3-Cy
clinA2sig
nalin
gpathway
[84]
PUMA
LAmiR-125bdirectlyrepressesP
umaan
apop
tosis
regulator
inp53pathway
[79]
BAK1
CDC2
5C
PPP1CA
PPP
2CAP
RKRA
TD
GTP5
3ZA
C1LA
miR-125bdirectlyrepressesa
poptosisregulatorssuchas
BAK1
CDC2
5CP
PP1C
AP
RKRA
and
TP53AZ
AC1in
p53pathway
[79]
PPP2
CART
-PCR
LA
miR-125bdirectlyinhibitsapop
tosis
bydo
wnregulatingPP
P2CA
expressio
n[85]
TDG
qRT-PC
RmiR-125bdirectlysupp
resses
thee
xpressionof
TDGin
p53pathway
[86]
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 373
Table1Targetso
foncogenicmiRNAs
andtheirroleincancer
cells
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-21
TPM1
WB
LAmiR-21d
ownregulates
TPM1inbreastcancer
andleadstoanchorage-independ
entg
rowth
[25]
E2F1
WB
RT-PCR
miR-21regulates
E2F1
byTG
F-120573sig
nalin
gpathway
[55]
TGFB
R2WB
RT-PCR
miR-21regulates
TGFB
R2by
TGF-120573sig
nalin
gpathway
[55]
TIMP3
WB
RT-PCR
TIMP3
isaM
MPinhibitormiR-21sup
pressesT
IMP3
byprom
otinginvasiv
enesso
fcancerc
ells
[56]
SERP
INB5
WB
LADow
nregulationof
PDCD
4andmaspin(SER
PINB5
)bymiR-21causestum
orogenesis
[57]
PDCD
4WB
RT-PCR
LA
miR-21d
ecreases
PDCD
4proteinam
ountsa
ndincreasesinvasion
[58]
FASCD
K6P
DCD
4CD
KN1A
FAM3C
HIPK3
PR
RG4AC
TA2BT
G2
BMPR
2SE
SN1IL6R
SOCS
5GLC
CI1APA
F1
SLC1
6A10SGK3
RP2
CFL
2
RT-PCR
M
Inhibitio
nof
miR-21inMCF
-7breastcancer
cells
show
sreduced
expressio
nof
p53tumor
supp
ressor
protein
FASCD
K6C
DKN
1AFAM3C
HIPK3
PRR
G4AC
TA2BT
G2BM
PR2
SESN
1IL6R
andtumor
supp
ressor
proteinProgrammed
CellD
eath
4(PDCD
4)area
lsodo
wnregulated
bymiR-21
[30]
FASTIMP3
WB
LADow
nregulationof
miR-21increases
FasligandandTIMP3
expressio
ntherebyitisinvolved
intheu
pregulationof
apop
tosis
[59]
CDC2
5AWB
RT-PCR
LA
miR-21sup
pressesC
dc25Aexpressio
nac
ellcycleregulator
[60]
RECK
RT-PCR
miR-21sup
pressesR
ECKandinhibitorsof
MMPs
andthus
prom
otes
invasiv
enesso
fcancerc
ells
[56]
MTA
PLA
Apop
totic
functio
nof
MTA
Pissupp
ressed
bymiR-21
[61]
RECK
WB
RT-PCR
LA
RECK
regu
latingthem
embrane-anchored
protease
isdo
wnregu
lated
bymiR-21
[62]
PDCD
4WB
LAmiR-21p
ositivelyregulates
PDCD
4throug
hinteractionwith
ovarianste
roidsa
ndTG
F-120573
signalin
gpathway
[55]
BTG2
EGFPW
BFluo
rescence
Intensity
Expressio
nof
miR-21leads
tolowe
rlevelof
BTG2which
isac
ellcycleregu
latora
ndtumor
supp
ressorItcauseslosso
fcon
trolonG1toSph
asetransition
[63]
PTEN
LAmiR-21regulates
MMP-2expressio
nin
cardiacfi
brob
lasts
oftheinfarctzone
viaP
TENpathway
[64]
BTG2
WB
RT-PCR
LA
miR-21d
irectlyinhibitsBT
G2which
isac
ellcycleinhibitorleading
toun
controlledDNA
synthesis
byactiv
ationof
forkhead
boxM1
[65]
PELI1
RT-PCR
LA
miR-21o
verexpressioninhibitsPeli1
viak
appaBsig
nalling
[66]
PDCD
4WB
Deregulationof
miR-21b
yvinylcarbam
ateind
uces
mou
selung
tumorigenesisby
downregulating
PDCD
4[67]
BIG-H
3LA
miR-21targetsbig-h3Itcou
ldbe
indu
cedby
TGF-120573anditregu
latesT
GF-120573sig
nalling
positively
andnegativ
ely
[68]
BIM
WB
LABima
apop
totic
activ
atorisc
ollectively
targeted
bymiR-21miR-24andmiR-221
andits
actio
nis
supp
ressed
[69]
HER
2WB
RT-PCR
miR-21u
pregulates
human
epidermalgrow
thfactor
receptor
2andactsas
oncogene
[34]
MSH
2SM
AD7
WB
RT-PCR
LA
miR-21d
ownregulates
MSH
2andSM
AD7in
TGF-120573pathway
andcauses
breastcancer
progression
[33]
374 Disease Markers
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-10b
HOXD
10WB
LAOverexpressionof
miR-10b
indu
cesrob
ustinvasionandmetastasis
bydo
wnregu
latin
gtransla
tionof
homeobo
xD10
[37]
KLF4
WB
RT-PCR
LA
miR-10b
redu
cese
ndogenou
sKLF
4protein
atum
orsupp
ressor
[70]
SFRS
1WB
RT-PCR
LA
miR-10b
supp
resses
SFRS
1(SR
-family
splicingfactor)a
ndcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
SFRS
10LA
miR-10b
supp
resses
SFRS
10andcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
miR-125a-5p
ERBB
2WB
RT-PCR
LA
Expressio
nof
either
miR-125ao
rmiR-125bresults
insupp
ressionof
ERBB
2andER
BB3
(oncogenefam
ily)a
ndexhibitsredu
cedmigratio
nandinvasio
ncapacitie
s[72]
TP53
WB
RT-PCR
LA
miR-125binhibitsp53gene
transla
tionally
andtranscrip
tionally
andtheisomer
ofmiRNA-
125
(miRNA-
125a)translationally
arrests
mRN
Aof
thep
53a
tumor
supp
ressor
gene
[44]
HuR
WB
RT-PCR
LA
HuR
isas
tressindu
cedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
Expressio
nof
miR-125as
ignificantly
redu
cesH
uR
[73]
ARID3B
WB
ARID3B
isoverexpressedin
human
ovariancancerO
verexpressionof
miR-125aind
uces
conversio
nof
high
lyinvasiv
eovaria
ncancer
cells
from
amesenchym
alto
anepith
elial
morph
ologyandsupp
resses
cancer
bynegativ
elyregu
latin
gEM
T[74]
miR-125b
BAK1
WB
RT-PCR
LAM
Overexpressionof
miR-125bstimulates
androgen-in
depend
entg
rowth
ofprostatecancer
bydo
wnregu
latin
gBa
k1(proapop
totic
regu
lator)
[75]
CDK6
CDC2
5AWB
miR-125boverexpressio
ndecreasesC
DK6
andCD
C25A
expressio
nItregu
latesp
roliferationof
U251g
liomas
tem
cells
throug
hcellcycle
regu
latedproteins
CDK6
andCD
C25A
[76]
SMO
WB
RT-PCR
LA
Dow
nregulationof
miR-125ballowsh
ighlevelsof
Hedgeho
g-depend
entgenee
xpressionleading
totumor
cellproliferatio
nby
allowingHedgeho
gsig
nalling
[77]
ST18
WB
BranchedL
AD
NA
Prob
eAssay
ST18
isdo
wnregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
DICER
1WB
LAB
ranchedDNA
Prob
eAssay
DICER
1isd
ownregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
TP53IN
P1LA
miR-125bdirectlyrepressesT
P53INP1anapop
tosis
regulator
inp53pathway
[79]
BMF
WB
RT-PCR
LA
overexpressio
nof
miR-125bredu
cese
xpressionof
cellapop
tosis
related
proteinBc
l-2mod
ifying
factor
(Bmf)bytargetingBM
F[80]
BMPR
1BLA
miR-125btargetsB
MPR
1Bandcauses
riskin
breastcancer
pathogenesis
[81]
HuR
WB
HuR
isas
tress-in
ducedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
miR-125atargetsHuR
[82]
120581B-Ra
s2LA
miR-125bdo
wnregulates
kapp
aB-Ras2gene
expressio
n[83]
E2F3
WB
RT-PCR
LA
E2F3anindu
cero
fbladd
ercancerissup
pressedby
miR-125batproteinlevelthrou
ghregulation
ofG1S
transitionthroug
htheE
2F3-Cy
clinA2sig
nalin
gpathway
[84]
PUMA
LAmiR-125bdirectlyrepressesP
umaan
apop
tosis
regulator
inp53pathway
[79]
BAK1
CDC2
5C
PPP1CA
PPP
2CAP
RKRA
TD
GTP5
3ZA
C1LA
miR-125bdirectlyrepressesa
poptosisregulatorssuchas
BAK1
CDC2
5CP
PP1C
AP
RKRA
and
TP53AZ
AC1in
p53pathway
[79]
PPP2
CART
-PCR
LA
miR-125bdirectlyinhibitsapop
tosis
bydo
wnregulatingPP
P2CA
expressio
n[85]
TDG
qRT-PC
RmiR-125bdirectlysupp
resses
thee
xpressionof
TDGin
p53pathway
[86]
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
374 Disease Markers
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-10b
HOXD
10WB
LAOverexpressionof
miR-10b
indu
cesrob
ustinvasionandmetastasis
bydo
wnregu
latin
gtransla
tionof
homeobo
xD10
[37]
KLF4
WB
RT-PCR
LA
miR-10b
redu
cese
ndogenou
sKLF
4protein
atum
orsupp
ressor
[70]
SFRS
1WB
RT-PCR
LA
miR-10b
supp
resses
SFRS
1(SR
-family
splicingfactor)a
ndcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
SFRS
10LA
miR-10b
supp
resses
SFRS
10andcauses
migratio
ninvasio
nandin
vivo
metastasis
ofneurob
lasto
mac
ells
[71]
miR-125a-5p
ERBB
2WB
RT-PCR
LA
Expressio
nof
either
miR-125ao
rmiR-125bresults
insupp
ressionof
ERBB
2andER
BB3
(oncogenefam
ily)a
ndexhibitsredu
cedmigratio
nandinvasio
ncapacitie
s[72]
TP53
WB
RT-PCR
LA
miR-125binhibitsp53gene
transla
tionally
andtranscrip
tionally
andtheisomer
ofmiRNA-
125
(miRNA-
125a)translationally
arrests
mRN
Aof
thep
53a
tumor
supp
ressor
gene
[44]
HuR
WB
RT-PCR
LA
HuR
isas
tressindu
cedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
Expressio
nof
miR-125as
ignificantly
redu
cesH
uR
[73]
ARID3B
WB
ARID3B
isoverexpressedin
human
ovariancancerO
verexpressionof
miR-125aind
uces
conversio
nof
high
lyinvasiv
eovaria
ncancer
cells
from
amesenchym
alto
anepith
elial
morph
ologyandsupp
resses
cancer
bynegativ
elyregu
latin
gEM
T[74]
miR-125b
BAK1
WB
RT-PCR
LAM
Overexpressionof
miR-125bstimulates
androgen-in
depend
entg
rowth
ofprostatecancer
bydo
wnregu
latin
gBa
k1(proapop
totic
regu
lator)
[75]
CDK6
CDC2
5AWB
miR-125boverexpressio
ndecreasesC
DK6
andCD
C25A
expressio
nItregu
latesp
roliferationof
U251g
liomas
tem
cells
throug
hcellcycle
regu
latedproteins
CDK6
andCD
C25A
[76]
SMO
WB
RT-PCR
LA
Dow
nregulationof
miR-125ballowsh
ighlevelsof
Hedgeho
g-depend
entgenee
xpressionleading
totumor
cellproliferatio
nby
allowingHedgeho
gsig
nalling
[77]
ST18
WB
BranchedL
AD
NA
Prob
eAssay
ST18
isdo
wnregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
DICER
1WB
LAB
ranchedDNA
Prob
eAssay
DICER
1isd
ownregulated
inDS-AMKL
patie
ntsh
avinghigh
expressio
nof
miR-125bthus
miR-125b-2actsas
apositive
regu
lator
ofmegakaryopo
iesis
[78]
TP53IN
P1LA
miR-125bdirectlyrepressesT
P53INP1anapop
tosis
regulator
inp53pathway
[79]
BMF
WB
RT-PCR
LA
overexpressio
nof
miR-125bredu
cese
xpressionof
cellapop
tosis
related
proteinBc
l-2mod
ifying
factor
(Bmf)bytargetingBM
F[80]
BMPR
1BLA
miR-125btargetsB
MPR
1Bandcauses
riskin
breastcancer
pathogenesis
[81]
HuR
WB
HuR
isas
tress-in
ducedRN
Abind
ingproteinhaving
over
expressio
nin
different
cancers
miR-125atargetsHuR
[82]
120581B-Ra
s2LA
miR-125bdo
wnregulates
kapp
aB-Ras2gene
expressio
n[83]
E2F3
WB
RT-PCR
LA
E2F3anindu
cero
fbladd
ercancerissup
pressedby
miR-125batproteinlevelthrou
ghregulation
ofG1S
transitionthroug
htheE
2F3-Cy
clinA2sig
nalin
gpathway
[84]
PUMA
LAmiR-125bdirectlyrepressesP
umaan
apop
tosis
regulator
inp53pathway
[79]
BAK1
CDC2
5C
PPP1CA
PPP
2CAP
RKRA
TD
GTP5
3ZA
C1LA
miR-125bdirectlyrepressesa
poptosisregulatorssuchas
BAK1
CDC2
5CP
PP1C
AP
RKRA
and
TP53AZ
AC1in
p53pathway
[79]
PPP2
CART
-PCR
LA
miR-125bdirectlyinhibitsapop
tosis
bydo
wnregulatingPP
P2CA
expressio
n[85]
TDG
qRT-PC
RmiR-125bdirectlysupp
resses
thee
xpressionof
TDGin
p53pathway
[86]
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 375
Table1Con
tinued
miRNAs
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-155
CYR6
1WB
RT-PCR
LA
overexpressio
nof
miR-155
contrib
utes
topreecla
mpsiadevelopm
entb
ytargetingand
downregulatingangiogenicregu
latingfactor
CYR6
1[87]
FOXO
3WB
RT-PCR
LA
Inversec
orrelationbetweenmiR-155
andFO
XO3a
prevailsin
breastcancer
celllin
esandtumors
[47]
SOCS
1Im
mun
ofluo
rescence
RT-PCR
LA
Overexpressionof
miR-155
inbreastcancer
cells
leadstoconstitutivea
ctivationof
signal
transducer
andactiv
ator
oftranscrip
tion3(STA
T3)throu
ghtheJanus-activated
kinase
(JAK)
pathway
[88]
CCND1
LAmiR-155
redu
cese
ndogenou
sexpressionof
CCND1b
ysupp
ressingLu
c-CC
ND1
[89]
IKBK
EWB
RT-PCR
LA
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
FADD
WB
RT-PCR
miR-155
expressio
nisincreasedin
gastric
epith
elialcelllines
andgastric
mucosaltissues
miR-155
negativ
elyregulates
IkappaBkinase
epsilon
Sma-
andMad-rela
tedprotein2(SMAD2)
Fas-associated
deathdo
mainprotein
interle
ukin-8and
grow
th-rela
tedon
cogene-alpha
[90]
JARID2
WB
RT-PCR
LA
overexpressio
nof
miR-155
decreaseslevels
ofendo
geno
usJARID2cellcycle
regu
latorm
RNAand
causes
tumor
[91]
CEBP
BWB
RT-PCR
LA
EBP120573
transla
tionissupp
ressed
bymiR-155
throug
hCEB
P120573in
tumor-activated
mon
ocytes
and
redu
cestum
orprogression
[92]
SHIP
WB
RT-PCR
LA
miR-155
decreasesS
HIP1expressionas
aresulto
fautocrin
estim
ulationby
proinfl
ammatory
cytokine
tumor
necrosisfactor
alph
a(TN
F-120572)
[93]
TAM
WB
RT-PCR
miR-155
downregulates
TAM
andcauses
spon
taneou
sbreastcancerd
evelo
pment
[94]
VHL
WB
RT-PCR
miR155do
wnregulates
VHLtumor
supp
ressor
viap
romotingHIF
transcrip
tionfactorsa
ndangiogenesis
[95]
p53
WB
RT-PCR
LA
miR155do
wnregulates
p53andprom
otes
cellproliferatio
n[52]
VHL
WB
RT-PCR
LA
miR155do
wnregulates
VHLandprom
otes
lymph
node
metastasis
andpo
orprogno
sisas
wellas
triple-negativetum
orin
breastcancer
[54]
CDC7
3WB
RT-PCR
LA
miR155negativ
elyregu
latesC
DC7
3which
inturn
positively
regu
lates120573
-catenincyclin
D1and
c-MYC
andincreasesc
ellviability
[53]
ZNF6
52WB
RT-PCR
miR155do
wnregulates
ZNF6
52causingexpressio
nof
TGFB
1TG
FB2TG
FBR2
EGFR
SMAD2
andVIM
resulting
inincreasedcellinvasio
nandmetastasis
[96]
MMP2
MMP9
VEG
FWB
RT-PCR
miR155causes
upregulationof
MMP2
MMP9
and
VEG
Fresulting
inincreasedtumor
size
[97]
HK2
WB
RT-PCR
LA
miR155up
regu
lates
hk2by
activ
atingsig
naltransdu
cera
ndactiv
ator
oftranscrip
tion3(STA
T3)
andtargetingCEB
P120573
[48]
WB
Western
blotR
T-PC
Rrealtim
e-PC
RLA
luciferase
AssayM
microarray
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
376 Disease Markers
hormone related gene ER-120572 (estrogen receptor alpha) [98]ER-120572 is important in estrogen related growth and its expres-sion level is marked in prognosis of ER-120572 positive breastcancer cells [99] There are two miR-206 binding sites withinthe 31015840-UTR of human ER-120572 and miR-206 downregulatesER-120572 mRNA and protein expression in breast cancer cells[99] The expression levels of miR-206 have been shownto be much higher in ER-120572 negative than in ER-120572 positivebreast cancer tumors and that the more ER-120572 positive cellspresent in the tumor the less miR-206 expression seensuggesting that miR-206 is a key factor for the regulationof ER-120572 expression in the development and progression ofhuman breast cancer [99] There was decreased miR-206expression in ER-120572 positive human breast cancer tissuesand that introduction of miR-206 into estrogen dependentbreast cancer cells inhibited cell growth [98]Thus it appearsthat miR-206 could be a novel candidate for ER120572-specificendocrine therapy in breast cancer Esterogen involves inbreast cancer cell proliferation by activating ER120572 receptorIt is recorded that around 70 of breast cancers are ERpositive Inhibitors of ER120572 such as tamoxifen and aromataseare widely used for treatment of ER120572 positive breast cancerpatients but during the treatment they are likely to prone forendocrine resistance [100] Hence there is a need for geneticlevel modifiers of ER120572 expression which could be achieved bymiRNAs In the mammary epithelium it was seen that miR-206 down regulates Tachykinin1 andGATA3 genes which areknown as breast cancer markers [101] The down regulationof miR-206 could lead to cancer progression by up regulatingexpression of cyclin D2 [102]
222 miR-31 miR-31 preventsmetastasis at multiple steps byinhibiting the expression of prometastatic genes Introduc-tion of miR-31 in metastatic breast cancer cells suppressedmetastasis-related functions like motility invasion and resis-tance to anoikis in vitro and metastasis in vivo [103] Theability of miR-31 to inhibit metastasis was attributable toinhibiting expression of RhoA The miR-31-mediated repres-sion of RhoA affected both local invasion and early postin-travasation events It was suggested that the full spectrum ofmiR-31rsquos effects on metastasis is elicited via the coordinaterepression of multiple targets [103] The expression of SATB2gene is associated with increased tumor cell migration andinvasion and miR-31 acts as a tumor suppressor by directlybinding to the 31015840UTR of SATB2 mRNA [104] miR-31 alsodirectly regulates FOXP3 by binding to the 31015840UTR of FOXP3mRNA [105] miR-31-P directly represses Dicer in MCF-7breast cancer cells [106] It has recently shown that miR-31expression inhibits the oncogenic NF-120581B pathway and thusit acts as a tumor suppressor miRNA [107]
223 miR-146 Breast cancer metastasis suppressor 1(BRMS1) regulates expression of multiple genes linked tometastasis including osteopontin (OPN) urokinase-typeplasminogen activator (uPA) epidermal growth factor recep-tor (EGFR) fascin and connexins [108] miR-146 is moreabundantly expressed in BRMS1-expressing cells miR-146a
and miR-146b are distinct genes encoded on chromosomes5q33 and 10q24 respectivelyThemature products have simi-lar targets because they differ only by twonucleotides near the31015840 end [108] miR-146a negatively regulates IFN pathway bytargeting the IFN regulatory factor 5 and STAT1 [109] miR-146a and miR-146b have been shown to inhibit both migra-tion and invasion suggesting that they play a role in metas-tasis [110] Since miR-146a andmiR-146b both reduce epider-mal growth factor receptor (EGFR) expression and suppressmetastasis [110] it is suggested that these miRNAs could havea therapeutic potential to suppress breast cancer metastasis
224 miR-91 miR-91 is also known as miR-17-5p locatedon chromosome 13q31 a genomic region that undergoesloss of heterozygosity in breast cancer [111] The oncogeneamplified in breast cancer (AIB1) is a direct target of miR-17-5p The protein encoded by the AIB1 gene is a steroidreceptor coactivator that enhances the transcriptional activityof ER120572 and E2F1 miR-17-5p represses the translation of AIB1mRNA thereby inhibiting the function of E2F1 and ER120572genes Down regulation of AIB1 by miR-17-5p results in sup-pression of estrogen stimulated proliferation and estrogenERindependent breast cancer cell proliferation [112] In breastcancer cells cyclin D1 (CCND1) which is over expressedin cancers was identified as a direct target of miR-17-5p Itwas shown that miR-17-5p inhibits the proliferation of breastcancer cells by suppressing cyclin D1 protein [113] Thereis higher expression of miR-17-5p in the noninvasive breastcancer cell lines (BT474 MCF7 MDA-MB-468 and T-47D)compared with highly invasive lines (MDA-MB-231 Hs578Tand SKBR3) [114] The detailed information about tumorsuppressor miRNAs their target genes and their regulationin breast cancer is listed in Table 2
23 OncogenicTumor Suppressor miRNAs
231 miR-205 The miRNA profiling data indicated thatbreast cancers without vascular invasion had high levelexpression of miR-205 compared to breast cancers withvascular invasion miR-205 was downregulated in breasttumor tissues as well as breast cancer cell lines in contrastmiR-205 was highly expressed in normal breast tissues and inthe nonmalignant breast epithelial cell line MCF-10A [72125] Cell proliferation was inhibited in MCF-7 cells whenthey were transfected and over-expressed with miR-205vector Over-expression of miR-205 allowed cells to grow onsoft agar very slowly indicating the reduced anchorage-inde-pendent growth The in vivo metastasis assays showed thatmiR-205 suppressed invasiveness in human breast cancercells (MDA-MB-231) [72]miR-205 targets and affects expres-sion of ErbB3 and VEGFA genes which are involved intumorogenecity [72 125] ErbB3 is amember of ErbB tyrosinekinase receptor family which is frequently overexpressed inbreast cancer There are correlations between ErbB3 expres-sion levels and tumorigenesis in breast cancer [126 127]VEGF-A is a key regulator of angiogenesis and it plays a keyrole particularly in tumormetastasismiR-205 directly targets
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 377
Table2Targetso
ftum
orsupp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esSh
ortd
escriptio
nRe
ferences
miR-206
ESR1
WB
RT-PCR
LA
miR-206
decreasese
strogenreceptor
(ER)alph
a1expressio
nandincreasese
xpressionof
hERa
lpha2It
also
causes
posttranscriptio
nalregulationof
ERalph
ainbreastcancer
[98]
ESR1
WB
RT-PCR
LA
miR-206
directlytargetsE
Ralpha
in31015840UTR
repo
rter
assays
[115]
MET
LAEx
pressio
nof
c-Metproteinisdo
wnregu
latedby
miR-206m
iR-206
expressio
nin
tumor
cells
show
sdelay
edgrow
thand
itcouldfunctio
nas
apotenttum
orsupp
ressor
inc-Met-overexpressingtumors
[116]
NOTC
H3
WB
RT-PCR
LA
miR-206
actsas
oncogene
aswellastum
orsupp
ressor
geneItind
uces
apop
totic
celldeathandblocks
antia
poptoticactiv
ityby
targetingNotch3
[117]
CyclinD
2WB
RT-PCR
LA
miR-206
targetstwobind
ingsites
inthe31015840UTR
ofcyclinD2mRN
Aandsupp
resses
cellproliferatio
nandcolony
form
ationatG1S
transition
[102]
miR-31
RHOA
WB
RT-PCR
LA
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis
prom
otinggenesR
hoA
[103]
FIH
WB
RT-PCR
LA
miR-31targetsFIHa
hypo
xia-indu
ciblefactor(HIF)regulatoryfactorthatinhibitsthea
bilityof
HIF
toactasa
transcrip
tionalregulator
undern
ormoxiccond
ition
s[118]
ITGA5RD
XMMP16fzd3M
PRIP
LAImmun
oBlot
miR-31expresses
inversely
with
metastasis
inhu
man
breastcancer
patie
ntsv
iasupp
ressionof
metastasis-promotinggenesF
zd3ITGA5MMP16MPR
IPand
RDX
[103]
SATB
2WB
RT-PCR
LA
SATB
2increasestum
orcellmigratio
nandinvasio
nwhilemiR-31isd
ownregu
lated
[104]
miR-146
CXCR
4LA
miR-146
aexpressiondecreasesC
XCR4
andinhibitsproliferatio
ndifferentiatio
nandmaturationas
well
asMKcolony
form
ation
[119]
CCNA2PA
2G4
BRCA
1M
miR-146
adecreases
expressio
nof
cellcycle
related
genes
[120]
IRF-5ST
AT1
WB
LAmiR-146
aexpressionredu
cesind
uctio
nof
type
IIFN
inthep
eripheralblood
mon
onuclear
cells
(PBM
Cs)
[109]
FADD
WB
LAmiR-146
aexpressionim
pairs
both
activ
ator
proteinAP-1a
ctivity
andinterle
ukin-2
prod
uctio
nindu
ced
byTC
Rengagement
[121]
MCP
-2EL
ISAL
A
miR-146
amaintains
HIV-m
ediatedchronicinfl
ammationof
brainby
decreasin
gthelevelof
MCP
2[122]
TBP
WB
LAmiR-146
atargetsTB
Pin
Hun
tington
diseasep
atho
genesis
[123]
MMP16
LAmiR-146
bincreasesg
liomac
ellm
igratio
nandinvasio
nviaM
MPs
[124]
miR-91
AIB1
WB
miR-91represses
transla
tionof
AIB1
[112]
CCND1
RT-PCR
miR-91inh
ibits
breastcancer
cellproliferatio
nby
targetingcyclinD1
[113]
E2F1
RT-PCR
miR-91d
ownregulates
thefun
ctionof
E2F1
byinhibitin
gAIB1
[112]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
378 Disease Markers
VEGF-A by interacting with the putative miR-205 bindingsite in its 31015840-UTRTherefore miR-205 can negatively regulatethe expression of ErbB3 and VEGF-A [72] and the resultssuggested that miR-205 may function as a tumor suppressorin breast cancer miR-205 can act as either tumor suppressoror oncogene and that is context-dependent [127]The expres-sion profile and the function of miRNAs vary in differentbreast cancer phenotypes (positive and negative ErbB2) Inaddition the patientsrsquo history such as age tumor virulencestage of tumor node status tumor proliferation index andother physical and chemical parameters may influence theactivity of miRNAs [43 72 128ndash130]
232 miR-27 miR-27a was upregulated in breast cancer andit appears to target the transcriptional cofactor zinc fingerand BTB domain containing 10 (ZBTB10) [131] ZBTB10represses the specificity protein 1 (SP1) transcription factorThe SP1 transcription factor is over expressed inmany cancersand plays a role in the G0-G1 to S phase progression in breastcancer cells [132] So it implies if expression of ZBTB10 isreduced miR-27a indirectly up regulates SP1 thus increasingS phase progression and functions as an oncogene miR-27a can regulate E2-responsiveness in MCF-7 cells throughsuppression of ZBTB10 thereby enhancing expression of ER[131] miR-27a also suppresses the cdc2cyclin B inhibitorMyt-1 in MDA-MB-231 cells and thereby facilitates breastcancer cell proliferationThus the oncogenic activity of miR-27a in MDA-MB-231 cells is due to suppression of expressionof ZBTB10 and Myt-1 proteins [132] It was hypothesizedthat miR-27 indirectly regulates the SP family protein withthe up regulation of the zinc finger ZBTB10 gene a putativeSP repressor [133] miR-27b a homologous to miR-27asharing 20 out of 21 nucleotides is potentially associatedwith cytochrome p4501b1 (CYP1B1) which is known tocatalyze the metabolism of certain procarcinogens and isover-expressed in a wide range of cancers [134 135] Inmost patients the expression level of miR-27b was found tobe decreased in cancerous tissues accompanied by a highlevel of CYP1B1 protein A significant inverse associationwas observed between the expression levels of miR-27b andCYP1B1 protein Thus the decreased expression of miR-27bwould be one of the causes of high expression of CYP1B1protein in cancerous tissues thus assuming the role for miR-27b as a tumor suppressor gene [135] Moreover it is evidentthat the HER2neu (ERBB2) EGF and TNFA promotemiR-23 and miR-27b expression (a prooncogenic factor)through the AKTNF-120581B signaling cascade [136] Thus theexpression and the function of miR-27 are likely to dependon the internal and external context of breast cancer andits phenotype As we mentioned earlier the patientsrsquo historysuch as age tumor virulence stage of tumor node statustumor proliferation index and other physical and chemicalparameters may influence the activity of miRNAs that isto act as either tumor suppressor or oncogene [43 72 128ndash130] In Table 3 we provide more detailed information aboutmiR-205 and miR-27 their target genes and their regulationduring breast cancer progression
3 Breast Cancer Bone Metastasis
Bone metastases are common in patients with advancedbreast cancer Breast cancer cells preferentially select bone tometastasize since bone matrix is enriched with nutrients thatsupport breast cancer cell growth Once tumor cells embedwithin the bone environment increased bone remodelingand bone destruction are common and are likely to mediateenhanced tumor growth in bone [137] Metastatic cancercells show a distinct biological profile from that of primarytumor cells by enhanced survival motility invasiveness andresistance to chemotherapy [4] Understanding the biologicalcontrols that signal tumor cell metastasis to bone and investi-gating the extent to which metastatic cancer cells differ fromprimary tumor cells with respect to miRNAs expression canprovide valuable insight into growth of tumors in the bonemicroenvironment
There are several lines of evidence indicating the involve-ment of specific miRNAs in suppressing or promotingbreast cancer metastasis and in other cancers [138 139]miRNA signatures that regulate breast cancer metastasis canbe observed either by tumor suppression or oncogenesisThey could either be downregulated (miR-30a3134a125s200s203205206342) or upregulated (miR-10b21135a155221222224373520c) In turn they influence the expressionof various target genes positively or negatively Neverthelessthey are potentially considered to be therapeutic agents intumor therapy [140] Metastasis otherwise known as tumormigration is regulated by a number of miRNAs via director indirect pathways The differential expression of miRNAsbetween metastatic cell lines was compared with the primarytumor and there was a down regulation of expression ofmiR-22 in metastatic cancer cells [141 142] Expression ofthe tumor suppressor miRNAs such as let-7 and miR-22 wasalso consistently downregulated in metastatic cancer cellsThese miRNAsrsquo target genes were found to be oncogenes(ERBB3 CDC25C and EVI-1T) Due to down regulationof miRNAs there were higher levels of expression of thesesignaling proteins in metastatic cells [141] Expression oftumor suppressor miRNAs such as let-7 miR-335 miR-126andmiR-206were downregulated [143] resulting in increasedmetastatic potential via up regulation of genes involved inmotility and proliferation miR-335 miR-206 and miR-126were selectively downregulated across a number of highlymetastatic human cell lines compared to the general tumorcell population and they had the ability to suppress metas-tasis of breast cancer cells to different organ sites miR-335suppresses metastasis and migration through targeting of theprogenitor cell transcription factor SOX4 and extracellularmatrix component tenascin C [143]
Up regulation of miRNAsrsquo expression inmetastases couldalso regress metastatic conditions Valastyan et al assessedthe therapeutic potential of acutely expressing miR-31 inalready-formed breast cancer metastases [142] Activation ofmiR-31 in established metastases elicits metastatic regressionand prolongs survival In contrast acute miR-31 expressionfails to affect primary mammary tumor growth [142] It ismost likely that a distinct role is played by specific miRNAand its expression of either down- or upregulation may not
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 379
Table3Targetso
foncogenictu
mor
supp
ressor
miRNAs
andtheirroleincancer
cells
miRNA
Targetgene
Techniqu
esShortd
escriptio
nRe
ferences
miR-205
ERBB
3WB
LAOvere
xpressionof
miR-205
downregulates
breasttumorsb
ydirectlytargetingHER
3receptor
and
inhibitsthea
ctivationof
thed
ownstre
ammediatorA
kt
[125]
ZEB1
LAmiR-205
isdo
wnregu
latedin
cells
thathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
TGF-120573Itd
ownregu
latesE
-cadherin
transcrip
tionalrepressorsZ
EB1and
enhances
tumor
metastasis
[144]
ZEB2
RT-PCR
miR-205
downregu
lates
E-cadh
erin
andredu
cesc
elllocom
otionandinvasio
nandexertsatum
orsupp
ressivee
ffect
[145]
ZEB2
LAmiR-205
downregu
latesc
ellsthathadun
dergon
eepithelialmesenchym
altransitionin
respon
seto
transfo
rminggrow
thfactor
(TGF-120573)Th
eydo
wnregu
lateE-cadh
erin
transcrip
tionalrepressorsZ
EB2and
enhancetum
ormetastasis
[144]
VEG
FALA
miR-205
inhibitscellproliferatio
nandanchorage-independ
entg
rowthasw
ellas
cellinvasio
nby
down
regu
latin
gtranscrip
tionof
VEG
FA
[72]
PRKC
EIm
mun
ostainingM
miR-205
downregu
lates
proteinkinase
Cepsilon
andredu
cesc
elllocom
otionandinvasio
nandexertsa
tumor
supp
ressivee
ffect
[145]
ZEB1Z
EB2
RT-PCR
WB
Mel-
8supp
resses
expressio
nof
miR-205
which
downregulates
ZEB1
andZE
B2resulting
inredu
ced
migratio
nandinvasio
n[128]
miR-27a
SP13
4WB
Expressio
nof
miR-27a
upregu
latesS
P1by
downregu
latin
gzinc
fingerZ
BTB10genea
putativ
eSp
repressortranscriptio
nally
[132]
FOXO
1WB
Severalm
iRNAs
areo
verexpressed
inendo
metria
lcancera
ndthey
repressF
OXO
1atum
orsupp
ressor
expressio
nresulting
inG1cellcyclearrestandcelldeath
[146]
FBXW
7WB
LAmiR-27a
overexpressio
nprom
otes
cellgrow
thby
supp
ressingFb
xw7forv
iraloncop
rotein
ST-in
duced
malignant
transfo
rmation
[147]
miR-27b
CYP1B1
WB
LAmiR-27b
supp
resses
expressio
nof
CYP1B1
inbreastcancer
andinhibitstumor
[135]
ST14
WB
RT-PCR
LA
ST14
redu
cesc
ellproliferation
migratio
nandinvasio
nmiR-27b
expressio
nincreasesd
uringcancer
progression
parallelin
gad
ecreaseinST
14expressio
nST
14redu
cesc
ellgrowth
throug
hits
effectson
cell
cycle
-related
proteins
[148]
MMP13
ELISAL
AIL-1120573-in
ducesa
ctivationof
signaltransdu
ctionpathwaysa
ssociatedwith
thee
xpressionof
MMP-13
and
downregulated
expressio
nof
miR-27b
inchon
drocytes
[149]
ZBTB
10WB
RT-PCR
miR-27do
wnregu
latese
xpressionof
ZBTB
10andincreasestum
orsiz
elymph
node
metastasisand
dista
ntmetastasis
[150]
WB
Western
BlotR
T-PC
RRe
alTime-PC
RLA
Luciferase
AssayM
Microarray
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
380 Disease Markers
have impact on both primary tumor growth and metastaticconditions Dicer is the key enzyme (ribonuclease) requiredfor the biogenesis of miRNAs and recent evidence indicatesthat Dicer may also be involved in tumorigenesis TheDicer RNA variants a and b contain a very long 31015840UTRwhereas c d and e variants contain very short 31015840UTRMultiple let-7-binding sites were found in Dicer codingas well as 31015840UTR Dicer mRNA expression is significantlycorrelated with the occurrence of distant metastases evenafter adjusting for other prognostic parameters In additionDicer mRNA expression has an independent prognosticvalue on metastatic disease in breast cancers [151] Thedownregulation of Dicer expression by let-7 miRNA may berelated to the metastatic spread of tumors [151] The recentresearches evolved with an understanding that there is a highprobability of interaction of three different genes (c-myccyclin D1 and Runx2) in bone metastasis Hence studyingthe regulation of these genes by miRNAs would be a valuableapproach for understanding of breast cancer progression tobone metastasis Since a number of miRNAs target c-myccyclin D1 and Runx2 genes in the following session we arefocusing on how these genes are regulated bymiRNAs duringbone metastasis
31 c-myc The signaling pathways or components playfundamental role in regulating cell morphology adhesionand motility Altering the signaling pathway could lead totumorigenesis and metastasis and miRNAs are found tobe involved in this process Rho-associated kinase (ROCK)is likely to enhance the metastatic propensity of breastcancer cells by stabilizing the actin cytoskeleton enhancingactin-myosin contraction and promoting the c-myc path-way including transcription of c-myc regulated miRNAsInhibition of ROCK-mediated signaling appears to be apromising and potentially specific approach to inhibit thebreast cancer metastases Further the expression of the c-myc regulated miR-17-92 cluster was elevated in metastaticbreast cancer cells compared with nonmetastatic cells [152]Furthermore blockade ofmiR-17 decreased breast cancer cellinvasionmigration in vitro and metastasis in vivo Togetherthese findings suggest that augmented ROCK signalingcontributes to breast cancer metastasis via indirectly upregulating miR-17 [152] Raf kinase inhibitory protein (RKIPalso PEBP1) a member of the evolutionarily conservedphosphatidylethanolamine-binding protein family has beenimplicated as a suppressor of metastatic progression RKIPrepresses invasion intravasation and bone metastasis ofbreast tumor cells in part through a signaling cascadeinvolving inhibition of MAPK myc and LIN28 leading toinduction of let-7 and down regulation of its targets [153]
c-myc protein (oncoprotein) can induce tumors with ahigh frequency and can induce massive programmed celldeath in most transgenic mouse model systems with greaterefficiency than other oncogenes [154] It was substantiatedwith evidence that miR-142 is involved in overexpressionof c-myc as it was translocated under the positive controlof the promoter region of miR-142 during a casual chro-mosomal rearrangement A mutation at the 31015840-UTR of
activation-induced cytidine deaminase (AID) mRNA thebinding site of miR-155 caused an increase in protein expres-sion of c-myc resulting in highMyc-Igc translocation a causeof cancer in Aicda mice [155] It was noticed that miR-24indirectly regulates c-myc via E2F2 thereby inhibiting cellproliferation [156] It has been shown that myc activatesexpression of miR-9 which gives cancer a metastatic status[157] The anaerobic metabolism of cancer cells (Warburgeffect) is the major cause for creating an acidic environ-ment to them and the cancer cellsrsquo energy source (ATP)is generated via glutaminolysis Mitochondrial glutaminasean important enzyme in this process is synthesized bytranscriptional repression of miR-23a and 23b by c-myc inhuman P-493 B lymphoma cells and PC3 prostate cancer cells[158] p53 can bind to its response element present inmiR-145promoter and can up regulatemiR-145 expression SincemiR-145 directly targets the 31015840UTR of c-myc gene it is likely thatp53 can regulate c-myc expression via miR-145 [159]
32 Cyclin D1 Cyclin D1 is widely accepted gene in thecontribution of oncogenesis of various cancerous cells andthe increased levels of cyclin D1 and c-myc in mouse breastcancer showed enhanced level of invasion and metastasis[154] Additionally cyclin D1 which upon genomic rear-rangement forms clonal lesions by overexpression in mul-tiple types of cancers [160] The highly conserved cluster ofmiR-17-92 acts as a negative regulator of cell proliferationThey sequentially are involved in the regulation of cyclin D1[113] and are regulated by c-myc via E-boxes in the promoter[161] whose dysregulation results in tumor A research on theregulatory feedback of miRNA 1720 suggested that miR-17-5pmiR-20a down regulates cyclin D1 in human breast cancerand their cell lines resulted in inhibition of serum-inducedS phase entry while in mammary epithelial cells cyclin D1after sufficient transcription induces the expression of miR-17 and miR-20 forming a cluster which results in a feedbackloop which in turn limits cell proliferation [113] In a genetarget prediction study it was notably identified that miR-193b in melanoma cell lines down regulates around 18 genesOne among these genes is cyclinD1This prediction can standas a basis for the study of the role of miR-193b in breastcancer [162] miR-200 is a family of miRNA that regulatescyclin D1 indirectly It was identified that in HeLa cells miR-200b binds to the 31015840UTR of RND3 directly and reduces itsexpression transcriptionally and translationally This in turnupregulated cyclin D1 protein expression and increased Sphase entry [163] Besides the up regulation of miR-98 andmiR-21 is known to decrease the expression of c-myc andE2F2 proteins through estrogen receptor 120572 [164] and theyact as tumor suppressors miR-503 is known to act as putativetumor suppressor and it targets cyclin D1 [165]
33 Runx2 There are several transcription factors beingimplicated with cancer progression The transcription factorRunx2 has an established role in cancers that metastasize tobone Runx2 is critical for regulation of genes that supportbone formation and is abnormally expressed in tumors thatmetastasize to bone The Runx genes comprise a family of
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 381
three closely related transcription factors Runx1 Runx2 andRunx3 which together with a common partner (CBF120573) formthe core binding factor (CBF) complex and bind DNA toeither activate or repress gene transcription While Runx1and Runx3 mutations are linked with leukemia and gastriccancer respectively [166 167] Runx2 has been proposedas one of the pivotal factors in the process of osteogenesisand metastasis in human malignancies including breastcancer [18ndash20] Runx2 can function as a metastasis-relatedoncoprotein in nonosseous cancer cells [168] In normalmammary epithelial cells Runx2 is expressed at low levelsbut it is expressed at high levels in metastatic breast cancercells [169 170] It is now widely recognized that breast cancercells preferentially metastasize to bone [50 171] It has shownthat inhibition of Runx2 activity abolishes the ability ofbreast cancer cells to form osteolytic lesions in vivo [50]Furthermore perturbation of Runx2 subnuclear localizationin breast cancer cells inhibits formation of osteolytic lesionsin bone in vivo
It was proposed that Runx2 activates cancer related genesin response to deregulated cell signaling pathways duringearly stages of breast cancer In recent years several miRNAshave been identified which can target Runx2 For examplemiR-155 and miR-355 directly target Runx2 [172 173] miR-155 targets the 31015840 untranslated region of multiple componentsof the BMP signaling cascade including SMAD1 SMAD5HIVEP2 CEBPB Runx2 and MYO10 miR-155 is expressedat elevated levels in human cancers including cancers of thelung breast colon and a subset of lymphoid malignancies[50] Runx2 expression can be downregulated by miRNAssuch as miR-133 miR-204211 [174 175] It is evident thatthe role of Runx2 is associated with breast cancer mediatedbone metastasis [50] Understanding miRNAs that regulateRunx2 activity supporting transformation of normal cellsto bone metastatic cancer cells would provide options fortherapeutic targets The role of miR-106b was determined inregulating Runx2 Cyclin D1 and myc via Wnt pathway with120573-catenin over expression Also the reduced expression levelof Cyclin D1 by miR-145 via ESR-1 endorsed the fact that thegene supports cell cycle progression [176] In silico validationidentified that miR-296 disrupts the EGFR signaling [177]Runx2 promotes differentiation of osteoblasts in wild typewith regulation of various cyclinsThe up regulation of Runx2during early G1 and down regulation prior to S phase indicateits role in cell proliferation [178ndash180]
Runx2 is functionally coupled with the cell motility inMDA-MB-231 cells This shows its role in the metastaticpotential of breast cancer cells with its capability to promoteinvasiveness and osteolytic property through a set of genesrequired for cell adhesion and motility by gain of functioneffect [180 181] Osteogenesis can be negatively regulatedby miR-204 and miR-211 in mesenchymal progenitor andBMSCs The repression of osteogenesis process could bedue to binding of miR-204211 to the 31015840-UTR of Runx2gene [174] It is reported that Runx2 stimulates Cyclin D1and reduces apoptosis by a synergy between Runx2 andc-myc [180 182] miR-204 205 217 133 and 135 weredownregulated in osteogenesis while miR-133 135 137 204205 217 and 218 were upregulated during chondrogenesis
and these miRNAs are recorded to regulate Runx2 [18] Theregulatorymechanism ofmiRNAs by Runx2 was identified inosteoblast differentiation with the formation of Runx2miR-3960miR-2861 feedback loop miR-3960 regulates the BMP2induced osteogenesis by targeting the Hoxa2 a repressor ofRunx2 [183] Eventhough it is evident that the role of Runx2is associated with breast cancer mediated bone metastasisfurther studies are needed to explore the miRNA expressionprofile targeting Runx2 in primary breast cancer growthand bone metastasis These studies suggest that the up- ordownregulation of one or more miRNAs would be a validapproach to alter expression of c-Myc Cyclin D1 and Runx2genes thereby prohibiting breast cancer progression to bonemetastasis
4 Conclusions and Future Perspectives
miRNAs are emerging group of ribonucleotides and studieson miRNAs and their expression are supportive for betterunderstanding of the network of genes and cellular pathwaysregulated by these miRNAs Recently the increased numbersof miRNAsrsquo analysis and their multiple target validation indifferent contexts of breast cancer are attaining a stronginsight in the field of miRNA based diagnosis and therapyfor breast cancer and bone metastasis There is differentialexpression and regulation of miRNAs associated with breastcancer growth and bonemetastasis which would help identi-fying them as possible biomarkers Potential therapies such asmiRNA silencing antisense blocking miRNAmodificationsor over-expression of miRNAs may be considered and theywould provide new opening avenues for more treatmentstrategies for patients with breast cancer and bonemetastasisBefore stepping into miRNA based diagnosis and treat-ment of breast cancer miRNA experimental expression tooland oligonucleotides strategies should be further developedmiRNA based therapy for breast cancer and bone metastasisrequires clinical evaluation of preferred miRNAsrsquo actionsat various levels Prior to systemic administration clinicaltrial is important to determine their exclusive targets sitespecificity and dose requirement The synthetic miRNAsrequire chemical modification(s) so that they can be stablein systemsrsquo fluid and can also be tissue specific Delivery ofmiRNAs to a specific tissuewould also require an appropriatenontoxic degradable delivery system Thus the maximizeddrug activity and the minimized side effects like toxicity andantigenicity should be considered for utilization of miRNAsin breast cancer therapy
Acknowledgment
This work was supported by grants from Indian Councilof Medical Research India (5132009-NCD-III 80102010-BMS to N Selvamurugan)
References
[1] ldquoMale breast cancer is a disease in which malignant (cancer)cells form in the tissues of the breastrdquo National Cancer Institute2013
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
382 Disease Markers
[2] ldquoWhat Are the Key Statistics About Breast Cancer in MenrdquoAmerican Cancer Society 2007
[3] C Desantis R Siegel P Bandi and A Jemal ldquoBreast cancerstatistics 2011rdquo CA Cancer Journal for Clinicians vol 61 no 6pp 409ndash418 2011
[4] Y Kang PM SiegelW Shu et al ldquoAmultigenic programmedi-ating breast cancer metastasis to bonerdquo Cancer Cell vol 3 no6 pp 537ndash549 2003
[5] S L Davis S G Eckhardt W A Messersmith and A JimenoldquoThe development of regorafenib and its current and potentialfuture role in cancer therapyrdquo Drugs of Today vol 49 pp 105ndash115 2013
[6] R Kumar J M Herman C L Wolfgang and L Zheng ldquoMulti-disciplinary management of pancreatic cancerrdquo Surgical Oncol-ogy Clinics of North America vol 22 pp 265ndash287 2013
[7] S Moore ldquoManaging treatment side effects in advanced breastcancerrdquo Seminars in Oncology Nursing vol 23 no 2 pp S23ndashS30 2007
[8] T A Buchholz ldquoRadiation therapy for early-stage breast cancerafter breast-conserving surgeryrdquo The New England Journal ofMedicine vol 360 no 1 pp 63ndash70 2009
[9] C K Zoon E Q Starker A MWilson M R Emmert-Buck SK Libutti and M A Tangrea ldquoCurrent molecular diagnosticsof breast cancer and the potential incorporation of microRNArdquoExpert Review of Molecular Diagnostics vol 9 no 5 pp 455ndash466 2009
[10] M V Iorio M Ferracin C-G Liu et al ldquoMicroRNA geneexpression deregulation in human breast cancerrdquo CancerResearch vol 65 no 16 pp 7065ndash7070 2005
[11] R I Gregory T P Chendrimada and R Shiekhattar ldquoMicroR-NA biogenesis isolation and characterization of the micropro-cessor complexrdquoMethods inMolecular Biology vol 342 pp 33ndash47 2006
[12] E P Murchison and G J Hannon ldquomiRNAs on the movemiRNA biogenesis and the RNAi machineryrdquo Current Opinionin Cell Biology vol 16 no 3 pp 223ndash229 2004
[13] A Grishok A E Pasquinelli D Conte et al ldquoGenes andmechanisms related to RNA interference regulate expression ofthe small temporal RNAs that control C elegans developmentaltimingrdquo Cell vol 106 no 1 pp 23ndash34 2001
[14] G D Bosse and M J Simard ldquoA new twist in the microRNApathway not dicer but argonaute is required for a microRNAproductionrdquo Cell Research vol 20 no 7 pp 735ndash737 2010
[15] D Cifuentes H Xue D W Taylor et al ldquoA novel miRNAprocessing pathway independent of dicer requires argonaute2catalytic activityrdquo Science vol 328 no 5986 pp 1694ndash16982010
[16] D Sayed and M Abdellatif ldquoMicroRNAs in development anddiseaserdquo Physiological Reviews vol 91 pp 827ndash887 2011
[17] T Kawamata M Yoda and Y Tomari ldquoMultilayer checkpointsfor microRNA authenticity during RISC assemblyrdquo EMBOReports vol 12 no 9 pp 944ndash949 2011
[18] J B Lian G S Stein A J VanWijnen et al ldquoMicroRNA controlof bone formation and homeostasisrdquo Nature Reviews Endocri-nology vol 8 no 4 pp 212ndash227 2012
[19] S Vimalraj and N Selvamurugan ldquoMicroRNAs synthesisgene regulation and osteoblast differentiationrdquoCurrent Issues inMolecular Biology vol 15 pp 7ndash18 2012
[20] A Moorthi S Vimalraj C Avani Z He N C Partridge andN Selvamurugan ldquoExpression of microRNA-30c and its target
genes in human osteoblastic cells by nano-bioglass ceramic-treatmentrdquo International Journal of Biological Macromoleculesvol 56 pp 181ndash185 2013
[21] J R Lytle T A Yario and J A Steitz ldquoTarget mRNAs arerepressed as efficiently by microRNA-binding sites in the 51015840UTR as in the 31015840 UTRrdquo Proceedings of the National Academyof Sciences of the United States of America vol 104 no 23 pp9667ndash9672 2007
[22] I Lee S S Ajay I Y Jong et al ldquoNew class of microRNA targetscontaining simultaneous 51015840-UTR and 31015840-UTR interaction sitesrdquoGenome Research vol 19 no 7 pp 1175ndash1183 2009
[23] J Brennecke A Stark R B Russell and S M Cohen ldquoPrinci-ples of microRNA-target recognitionrdquo PLoS Biology vol 3 no3 article e85 2005
[24] P Mestdagh S Lefever F Pattyn et al ldquoThe microRNA bodymap dissecting microRNA function through integrativegenomicsrdquo Nucleic Acids Research vol 39 no 20 article e1362011
[25] S Zhu M-L Si H Wu and Y-Y Mo ldquoMicroRNA-21 targetsthe tumor suppressor gene tropomyosin 1 (TPM1)rdquo Journal ofBiological Chemistry vol 282 no 19 pp 14328ndash14336 2007
[26] J Boyd J I Risinger RWWiseman B AMerrick J K Selkirkand J C Barrett ldquoRegulation ofmicrofilament organization andanchorage-independent growth by tropomyosin 1rdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 25 pp 11534ndash11538 1995
[27] LQi J Bart L P Tan et al ldquoExpression ofmiR-21 and its targets(PTEN PDCD4 TM1) in flat epithelial atypia of the breast inrelation to ductal carcinoma in situ and invasive carcinomardquoBMC Cancer vol 9 article 163 2009
[28] I Sansal and W R Sellers ldquoThe biology and clinical relevanceof the PTEN tumor suppressor pathwayrdquo Journal of ClinicalOncology vol 22 no 14 pp 2954ndash2963 2004
[29] H-S Yang A P Jansen R Nair et al ldquoA novel transformationsuppressor Pdcd4 inhibits AP-1 transactivation but not NF-120581Bor ODC transactivationrdquo Oncogene vol 20 no 6 pp 669ndash6762001
[30] L B Frankel N R Christoffersen A Jacobsen M Lindow AKrogh and A H Lund ldquoProgrammed cell death 4 (PDCD4)is an important functional target of the microRNA miR-21 inbreast cancer cellsrdquo Journal of Biological Chemistry vol 283 no2 pp 1026ndash1033 2008
[31] B Lankat-Buttgereit and R Goke ldquoThe tumour suppressorPdcd4 recent advances in the elucidation of function andregulationrdquo Biology of the Cell vol 101 no 6 pp 309ndash317 2009
[32] M Terao M Fratelli M Kurosaki et al ldquoInduction of miR-21by retinoic acid in estrogen receptor-positive breast carcinomacells biological correlates and molecular targetsrdquo Journal ofBiological Chemistry vol 286 no 5 pp 4027ndash4042 2011
[33] L Chen Y Li Y Fu et al ldquoRole of deregulated microRNAs inbreast cancer progression using FFPE tissuerdquo PLoS ONE vol 8Article ID e54213 2013
[34] A Ozgun B Karagoz O Bilgi T Tuncel H Baloglu and E GKandemir ldquoMicroRNA-21 as an indicator of aggressive pheno-type in breast cancerrdquo Onkologie vol 36 pp 115ndash118 2013
[35] L X Yan Q N Wu Y Zhang et al ldquoKnockdown of miR-21in human breast cancer cell lines inhibits proliferation in vitromigration and in vivo tumor growthrdquo Breast Cancer Researchvol 13 no 1 article R 2011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 383
[36] F Pichiorri D Palmieri L De Luca et al ldquoIn vivoNCL targetingaffects breast cancer aggressiveness through miRNA regula-tionrdquo Journal of Experimental Medicine vol 210 pp 951ndash9682013
[37] L Ma J Teruya-Feldstein and R A Weinberg ldquoTumourinvasion and metastasis initiated by microRNA-10b in breastcancerrdquo Nature vol 449 no 7163 pp 682ndash688 2007
[38] L Ma ldquoRole of miR-10b in breast cancer metastasisrdquo BreastCancer Research vol 12 no 5 article 210 2010
[39] M Negrini and G A Calin ldquoBreast cancer metastasis amicroRNA storyrdquo Breast Cancer Research vol 10 no 2 article303 2008
[40] F Biagioni N Bossel Ben-Moshe G Fontemaggi et al ldquomiR-10blowast a master inhibitor of the cell cycle is down-regulated inhuman breast tumoursrdquo EMBO Molecular Medicine vol 4 pp1214ndash1229 2012
[41] S A Ibrahim GW Yip C Stock et al ldquoTargeting of syndecan-1 by microRNA miR-10b promotes breast cancer cell motilityand invasiveness via aRho-GTPase- andE-cadherin-dependentmechanismrdquo International Journal of Cancer vol 131 pp E884ndashE896 2012
[42] G K Scott A Goga D Bhaumik C E Berger C S Sullivanand C C Benz ldquoCoordinate suppression of ERBB2 and ERBB3by enforced expression of micro-RNA miR-125a or miR-125brdquoJournal of Biological Chemistry vol 282 no 2 pp 1479ndash14862007
[43] M D Mattie C C Benz J Bowers et al ldquoOptimized high-throughput microRNA expression profiling provides novelbiomarker assessment of clinical prostate and breast cancerbiopsiesrdquoMolecular Cancer vol 5 article 24 2006
[44] Y Zhang J-S Gao X Tang et al ldquoMicroRNA 125a and itsregulation of the p53 tumor suppressor generdquo FEBS Letters vol583 no 22 pp 3725ndash3730 2009
[45] E OrsquoDay and A Lal ldquoMicroRNAs and their target genenetworks in breast cancerrdquo Breast Cancer Research vol 12 no2 article 201 2010
[46] W Kong H Yang L He et al ldquoMicroRNA-155 is regu-lated by the transforming growth factor 120573Smad pathway andcontributes to epithelial cell plasticity by targeting RhoArdquoMolecular and Cellular Biology vol 28 no 22 pp 6773ndash67842008
[47] WKong L HeM Coppola et al ldquoMicroRNA-155 regulates cellsurvival growth and chemosensitivity by targeting FOXO3a inbreast cancerrdquo Journal of Biological Chemistry vol 285 no 23pp 17869ndash17879 2010
[48] S Jiang L-F Zhang H-W Zhang et al ldquoA novel miR-155miR-143 cascade controls glycolysis by regulating hexokinase 2 inbreast cancer cellsrdquo EMBO Journal vol 31 no 8 pp 1985ndash19982012
[49] C Roth B Rack V Muller W Janni K Pantel andH Schwarzenbach ldquoCirculating microRNAs as blood-basedmarkers for patients with primary andmetastatic breast cancerrdquoBreast Cancer Research vol 12 no 6 article R90 2010
[50] Q Yin X Wang C Fewell et al ldquoMicroRNA miR-155 inhibitsBone Morphogenetic Protein (BMP) signaling and BMP-mediated Epstein-Barr virus reactivationrdquo Journal of Virologyvol 84 no 13 pp 6318ndash6327 2010
[51] C Zhang J Zhao and H Deng ldquo17120573-Estradiol up-regulatesmiR-155 expression and reduces TP53INP1 expression in MCF-7 breast cancer cellsrdquoMolecular and Cellular Biochemistry vol379 pp 201ndash211 2013
[52] J Liu Q Mao Y Liu X Hao S Zhang and J Zhang ldquoAnalysisofmiR-205 andmiR-155 expression in the blood of breast cancerpatientsrdquoChinese Journal of Cancer Research vol 25 pp 46ndash542013
[53] M I Rather M N Nagashri S S Swamy K S Gopinath andA Kumar ldquoOncogenic microRNA-155 down-regulates tumorsuppressor CDC73 and promotes oral squamous cell carcinomacell proliferation implications for cancer therapeuticsrdquo TheJournal of Biological Chemistry vol 288 pp 608ndash618 2013
[54] W Kong L He E J Richards et al ldquoUpregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL andis associated with poor prognosis and triple-negative breastcancerrdquo Oncogene 2013
[55] Q Pan X Luo and N Chegini ldquomicroRNA 21 response tohormonal therapies and regulatory function in leiomyomatransformed leiomyoma and leiomyosarcoma cellsrdquo MolecularHuman Reproduction vol 16 no 3 Article ID gap093 pp 215ndash227 2009
[56] G Gabriely T Wurdinger S Kesari et al ldquoMicroRNA 21 pro-motes glioma invasion by targeting matrix metalloproteinaseregulatorsrdquo Molecular and Cellular Biology vol 28 no 17 pp5369ndash5380 2008
[57] S Zhu H Wu F Wu D Nie S Sheng and Y-Y Mo ldquoMicroR-NA-21 targets tumor suppressor genes in invasion and metasta-sisrdquo Cell Research vol 18 no 3 pp 350ndash359 2008
[58] I A Asangani S A K Rasheed D A Nikolova et al ldquoMicroR-NA-21 (miR-21) post-transcriptionally downregulates tumorsuppressor Pdcd4 and stimulates invasion intravasation andmetastasis in colorectal cancerrdquo Oncogene vol 27 no 15 pp2128ndash2136 2008
[59] A Zhang Y Liu Y Shen Y Xu and X Li ldquoMiR-21 modulatescell apoptosis by targeting multiple genes in renal cell carci-nomardquo Urology vol 78 no 2 pp 474e13ndash474e19 2011
[60] P Wang F Zou Z Xiaodong et al ldquomicroRNA-21 negativelyregulates Cdc25A and cell cycle progression in colon cancercellsrdquo Cancer Research vol 69 no 20 pp 8157ndash8165 2009
[61] Y Chen W Liu T Chao et al ldquoMicroRNA-21 down-regulatesthe expression of tumor suppressor PDCD4 in human glioblas-toma cell T98Grdquo Cancer Letters vol 272 no 2 pp 197ndash2052008
[62] F Loayza-Puch Y Yoshida T Matsuzaki C Takahashi HKitayama and M Noda ldquoHypoxia and RAS-signaling path-ways converge on and cooperatively downregulate the RECKtumor-suppressor protein throughmicroRNAsrdquoOncogene vol29 no 18 pp 2638ndash2648 2010
[63] M Liu H Wu T Liu et al ldquoRegulation of the cell cycle geneBTG2 bymiR-21 in human laryngeal carcinomardquoCell Researchvol 19 no 7 pp 828ndash837 2009
[64] S Roy S Khanna S-R A Hussain et al ldquoMicroRNA expres-sion in response to murine myocardial infarction MiR-21 reg-ulates fibroblast metalloprotease-2 via phosphatase and tensinhomologuerdquo Cardiovascular Research vol 82 no 1 pp 21ndash292009
[65] G Song A D Sharma G R Roll et al ldquoMicroRNAs controlhepatocyte proliferation during liver regenerationrdquoHepatologyvol 51 no 5 pp 1735ndash1743 2010
[66] R T Marquez E Wendlandt C S Galle K Keck and A PMcCaffrey ldquoMicroRNA-21 is upregulated during the prolifera-tive phase of liver regeneration targets Pellino-1 and inhibitsNF-120581B signalingrdquo American Journal of Physiology vol 298 no4 pp G535ndashG541 2010
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
384 Disease Markers
[67] TMelkamu X Zhang J Tan Y Zeng and F Kassie ldquoAlterationof microRNA expression in vinyl carbamate-induced mouselung tumors and modulation by the chemopreventive agentindole-3-carbinolrdquo Carcinogenesis vol 31 no 2 pp 252ndash2582010
[68] C Liu B Li Y Cheng et al ldquoMiR-21 plays an important rolein radiation induced carcinogenesis in BALBc mice by directlytargeting the tumor suppressor gene Big-h3rdquo InternationalJournal of Biological Sciences vol 7 no 3 pp 347ndash363 2011
[69] S Hu M Huang P K Nguyen et al ldquoNovel MicroRNAprosurvival cocktail for improving engraftment and function ofcardiac progenitor cell transplantationrdquoCirculation vol 124 no11 pp S27ndashS34 2011
[70] Y Tian A Luo Y Cai et al ldquoMicroRNA-10b promotes migra-tion and invasion through KLF4 in human esophageal cancercell linesrdquo Journal of Biological Chemistry vol 285 no 11 pp7986ndash7994 2010
[71] S Meseguer G Mudduluru J M Escamilla H Allgayer andD Barettino ldquoMicroRNAs-10a and -10b contribute to retinoicacid-induced differentiation of neuroblastoma cells and targetthe alternative splicing regulatory factor SFRS1 (SF2ASF)rdquoJournal of Biological Chemistry vol 286 no 6 pp 4150ndash41642011
[72] H Wu S Zhu and Y-Y Mo ldquoSuppression of cell growth andinvasion by miR-205 in breast cancerrdquo Cell Research vol 19 no4 pp 439ndash448 2009
[73] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[74] K D Cowden Dahl R Dahl J N Kruichak and L G HudsonldquoThe epidermal growth factor receptor responsive miR-125arepresses mesenchymal morphology in ovarian cancer cellsrdquoNeoplasia vol 11 no 11 pp 1208ndash1215 2009
[75] X-B Shi L Xue J Yang et al ldquoAn androgen-regulated miRNAsuppresses Bak1 expression and induces androgen-independentgrowth of prostate cancer cellsrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 104 no50 pp 19983ndash19988 2007
[76] L Shi J Zhang T Pan et al ldquoMiR-125b is critical for the sup-pression of human U251 glioma stem cell proliferationrdquo BrainResearch vol 1312 pp 120ndash126 2010
[77] E Ferretti E De Smaele E Miele et al ldquoConcerted microRNAcontrol of Hedgehog signalling in cerebellar neuronal progeni-tor and tumour cellsrdquo EMBO Journal vol 27 no 19 pp 2616ndash2627 2008
[78] J-H Klusmann Z Li K Bohmer et al ldquomiR-125b-2 is a poten-tial oncomiR on human chromosome 21 in megakaryoblasticleukemiardquo Genes and Development vol 24 no 5 pp 478ndash4902010
[79] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[80] H-F Xia T-Z He C-M Liu et al ldquoMiR-125b expressionaffects the proliferation and apoptosis of human glioma cells bytargeting BmfrdquoCellular Physiology and Biochemistry vol 23 no4ndash6 pp 347ndash358 2009
[81] P Saeligtrom J Biesinger S M Li et al ldquoA risk variant in an miR-125b binding site in BMPR1B is associated with breast cancerpathogenesisrdquo Cancer Research vol 69 no 18 pp 7459ndash74652009
[82] X Guo YWu and R S Hartley ldquoMicroRNA-125a represses cellgrowth by targeting HuR in breast cancerrdquo RNA Biology vol 6no 5 pp 575ndash583 2009
[83] A J Murphy P M Guyre and P A Pioli ldquoEstradiol suppressesNF-120581B activation through coordinated regulation of let-7aand miR-125b in primary human macrophagesrdquo Journal ofImmunology vol 184 no 9 pp 5029ndash5037 2010
[84] L Huang J Luo Q Cai et al ldquoMicroRNA-125b suppresses thedevelopment of bladder cancer by targeting E2F3rdquo InternationalJournal of Cancer vol 128 no 8 pp 1758ndash1769 2011
[85] M Bousquet D Nguyen C Chen L Shields and H F LodishldquoMicroRNA-125b transforms myeloid cell lines by repressingmultiple mRNArdquo Haematologica vol 97 pp 1713ndash1721 2012
[86] M T N Le N Shyh-Chang S L Khaw et al ldquoConservedregulation of p53 network dosage by microRNA-125b occursthrough evolving miRNA-target gene pairsrdquo PLoS Genetics vol7 no 9 Article ID e1002242 2011
[87] Y Zhang Z Diao L Su et al ldquoMicroRNA-155 contributes topreeclampsia by down-regulating CYR61rdquo American Journal ofObstetrics and Gynecology vol 202 no 5 pp 466e1ndash466e72010
[88] S Jiang H-W Zhang M-H Lu et al ldquoMicroRNA-155 func-tions as an oncomiR in breast cancer by targeting the suppressorof cytokine signaling 1 generdquo Cancer Research vol 70 no 8 pp3119ndash3127 2010
[89] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[90] B Xiao Z Liu B-S Li et al ldquoInduction of microRNA-155 dur-ing helicobacter pylori infection and its negative regulatory rolein the inflammatory responserdquo Journal of Infectious Diseasesvol 200 no 6 pp 916ndash925 2009
[91] M T Bolisetty G Dy W Tam and K L Beemon ldquoReticu-loendotheliosis virus strain T induces miR-155 which targetsJARID2 and promotes cell survivalrdquo Journal of Virology vol 83no 23 pp 12009ndash12017 2009
[92] MHe Z Xu TDingD-MKuang and L Zheng ldquoMicroRNA-155 regulates inflammatory cytokine production in tumor-associated macrophages via targeting CEBP120573rdquo Chinese Journalof Cellular andMolecular Immunology vol 6 no 5 pp 343ndash3522009
[93] I M Pedersen D Otero E Kao et al ldquoOnco-miR-155 targetsSHIP1 to promote TNF120572-dependent growth of B cell lym-phomasrdquo EMBO Molecular Medicine vol 1 no 5 pp 288ndash2952009
[94] E Zonari F PucciM Saini et al ldquoA role formiR-155 in enablingtumor-infiltrating innate immune cells to mount effective anti-tumor responses in micerdquo Blood vol 122 pp 243ndash252 2013
[95] M F Czyzyk-Krzeska and X Zhang ldquoMiR-155 at the heart ofoncogenic pathwaysrdquo Oncogene 2013
[96] P M Neilsen J E Noll S Mattiske et al ldquoMutant p53 drivesinvasion in breast tumors through up-regulation of miR-155rdquoOncogene vol 32 pp 2992ndash3000 2012
[97] M M Hafez Z K Hassan A R Zekri et al ldquoMicroRNAs andmetastasis-related gene expression in Egyptian breast cancerpatientsrdquo Asian Pacific Journal of Cancer Prevention vol 13 pp591ndash598 2012
[98] B D Adams H Furneaux and B A White ldquoThe micro-ribonucleic acid (miRNA) miR-206 targets the human estro-gen receptor-120572 (ER120572) and represses ER120572 messenger RNAand protein expression in breast cancer cell linesrdquo MolecularEndocrinology vol 21 no 5 pp 1132ndash1147 2007
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 385
[99] N Kondo T Toyama H Sugiura Y Fujii and H YamashitaldquomiR-206 expression is down-regulated in estrogen receptor 120572-positive human breast cancerrdquo Cancer Research vol 68 no 13pp 5004ndash5008 2008
[100] R Garcıa-Becerra N Santos L Dıaz and J Camacho ldquoMech-anisms of resistance to endocrine therapy in breast cancerfocus on signaling pathways miRNAs and genetically basedresistancerdquo International Journal of Molecular Sciences vol 14pp 108ndash145 2012
[101] M J Lee K S Yoon K W Cho K S Kim and H S JungldquoExpression of miR-206 during the initiation of mammarygland developmentrdquoCell and Tissue Research vol 353 no 3 pp425ndash433 2013
[102] J Zhou Y Tian J Li et al ldquomiR-206 is down-regulated inbreast cancer and inhibits cell proliferation through the up-regulation of cyclinD2rdquo Biochemical and Biophysical ResearchCommunications vol 433 pp 207ndash212 2013
[103] S Valastyan F Reinhardt N Benaich et al ldquoA pleiotropicallyacting MicroRNA miR-31 inhibits breast cancer metastasisrdquoCell vol 137 no 6 pp 1032ndash1046 2009
[104] O Aprelikova X Yu J Palla et al ldquoThe role of miR-31 and itstarget gene SATB2 in cancer-associated fibroblastsrdquo Cell Cyclevol 9 no 21 pp 4387ndash4398 2010
[105] R Rouas H Fayyad-Kazan N El Zien et al ldquoHumannatural Treg microRNA signature role of microRNA-31 andmicroRNA-21 in FOXP3 expressionrdquo European Journal ofImmunology vol 39 no 6 pp 1608ndash1618 2009
[106] Y T Chan Y C Lin R J Lin et al ldquoConcordant and discordantregulation of target genes by miR-31 and its isoformsrdquo PLoSONE vol 8 Article ID e58169 2013
[107] C Korner I Keklikoglou C Bender A Worner E Mun-stermann and S Wiemann ldquoMicroRNA-31 sensitizes humanbreast cells to apoptosis by direct targeting of protein kinase Cepsilon (PKCepsilon)rdquoThe Journal of Biological Chemistry vol288 pp 8750ndash8761 2013
[108] D R Hurst M D Edmonds G K Scott C C Benz K SVaidya and D R Welch ldquoBreast cancer metastasis suppressor1 up-regulates miR-146 Which suppresses breast cancer metas-tasisrdquo Cancer Research vol 69 no 4 pp 1279ndash1283 2009
[109] Y Tang X Luo H Cui et al ldquoMicroRNA-146a contributes toabnormal activation of the type I interferon pathway in humanlupus by targeting the key signaling proteinsrdquo Arthritis andRheumatism vol 60 no 4 pp 1065ndash1075 2009
[110] D Bhaumik G K Scott S Schokrpur C K Patil J Campisiand C C Benz ldquoExpression of microRNA-146 suppresses NF-120581B activity with reduction of metastatic potential in breastcancer cellsrdquo Oncogene vol 27 no 42 pp 5643ndash5647 2008
[111] G Eiriksdottir G Johannesdottir S Ingvarsson et al ldquoMappingloss of heterozygosity at chromosome 13q loss at 13q12-q13 isassociated with breast turnout progression and poor prognosisrdquoEuropean Journal of Cancer vol 34 no 13 pp 2076ndash2081 1998
[112] A Hossain M T Kuo and G F Saunders ldquoMir-17-5p regulatesbreast cancer cell proliferation by inhibiting translation of AIB1mRNArdquoMolecular and Cellular Biology vol 26 no 21 pp 8191ndash8201 2006
[113] Z Yu C Wang M Wang et al ldquoA cyclin D1microRNA1720 regulatory feedback loop in control of breast cancer cellproliferationrdquo Journal of Cell Biology vol 182 no 3 pp 509ndash517 2008
[114] Z Yu N E Willmarth J Zhou et al ldquomicroRNA 1720 inhibitscellular invasion and tumor metastasis in breast cancer by
heterotypic signalingrdquo Proceedings of the National Academy ofSciences of the United States of America vol 107 no 18 pp 8231ndash8236 2010
[115] S-K Leivonen R Makela P Ostling et al ldquoProtein lysatemicroarray analysis to identify microRNAs regulating estrogenreceptor signaling in breast cancer cell linesrdquoOncogene vol 28no 44 pp 3926ndash3936 2009
[116] D Yan X D Dong X Chen et al ldquoMicroRNA-1206 targets c-met and inhibits rhabdomyosarcoma developmentrdquo Journal ofBiological Chemistry vol 284 no 43 pp 29596ndash29604 2009
[117] G Song Y Zhang and L Wang ldquoMicroRNA-206 targetsnotch3 activates apoptosis and inhibits tumor cell migrationand focus formationrdquo Journal of Biological Chemistry vol 284no 46 pp 31921ndash31927 2009
[118] C-J Liu M-M Tsai P-S Hung et al ldquomiR-31 ablates expres-sion of the HIF regulatory factor FIH to activate the HIFpathway in head and neck carcinomardquo Cancer Research vol 70no 4 pp 1635ndash1644 2010
[119] C Labbaye I SpinelloM T Quaranta et al ldquoA three-step path-way comprising PLZFmiR-146aCXCR4 controls megakary-opoiesisrdquo Nature Cell Biology vol 10 no 7 pp 788ndash801 2008
[120] C-H Hsieh C-S Rau S-F Jeng et al ldquoIdentification of thepotential target genes of microRNA-146a induced by PMAtreatment in human microvascular endothelial cellsrdquo Experi-mental Cell Research vol 316 no 7 pp 1119ndash1126 2010
[121] G Curtale F Citarella C Carissimi et al ldquoAn emergingplayer in the adaptive immune response MicroRNA-146a is amodulator of IL-2 expression and activation-induced cell deathin T lymphocytesrdquo Blood vol 115 no 2 pp 265ndash273 2010
[122] S Rom I Rom G Passiatore et al ldquoCCL8MCP-2 is a targetfor mir-146a in HIV-1-infected human microglial cellsrdquo FASEBJournal vol 24 no 7 pp 2292ndash2300 2010
[123] M Sinha J Ghose E Das and N P Bhattarcharyya ldquoAlteredmicroRNAs in STHdhQ111HdhQ111 cells miR-146a targetsTBPrdquo Biochemical and Biophysical Research Communicationsvol 396 no 3 pp 742ndash747 2010
[124] H Xia Y Qi S S Ng et al ldquomicroRNA-146b inhibits gliomacellmigration and invasion by targetingMMPsrdquoBrain Researchvol 1269 pp 158ndash165 2009
[125] M V Iorio P Casalini C Piovan et al ldquoMicroRNA-205regulates HER3 in human breast cancerrdquo Cancer Research vol69 no 6 pp 2195ndash2200 2009
[126] T Holbro R R Beerli F Maurer M Koziczak C F BarbasIII and N E Hynes ldquoThe ErbB2ErbB3 heterodimer functionsas an oncogenic unit ErbB2 requires ErbB3 to drive breasttumor cell proliferationrdquo Proceedings of the National Academyof Sciences of the United States of America vol 100 no 15 pp8933ndash8938 2003
[127] S B Greene J I Herschkowitz and J M Rosen ldquoThe upsand downs of miR-205 identifying the roles of miR-205 inmammary gland development and breast cancerrdquo RNA Biologyvol 7 no 3 2010
[128] J Y Lee M K Park J H Park et al ldquoLoss of the polycombprotein Mel-18 enhances the epithelial-mesenchymal transitionby ZEB1 and ZEB2 expression through the downregulation ofmiR-205 in breast cancerrdquo Oncogene 2013
[129] C Piovan D Palmieri G Di Leva et al ldquoOncosuppressiverole of p53-induced miR-205 in triple negative breast cancerrdquoMolecular Oncology vol 6 pp 458ndash472 2012
[130] M Tanic M Zajac G Gomez-Lopez J Benıtez and BMartınez-Delgado ldquoIntegration of BRCA1-mediated miRNA
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
386 Disease Markers
andmRNAprofiles revealsmicroRNA regulation of TRAF2 andNF120581B pathwayrdquo Breast Cancer Research and Treatment vol 134pp 41ndash51 2012
[131] X Li S U Mertens-Talcott S Zhang K Kim J Ball and SSafe ldquoMicroRNA-27a indirectly regulates estrogen receptor120572 expression and hormone responsiveness in MCF-7 breastcancer cellsrdquo Endocrinology vol 151 no 6 pp 2462ndash2473 2010
[132] S U Mertens-Talcott S Chintharlapalli X Li and S Safe ldquoTheoncogenic microRNA-27a targets genes that regulate specificityprotein transcription factors and the G2-M checkpoint inMDA-MB-231 breast cancer cellsrdquo Cancer Research vol 67 no22 pp 11001ndash11011 2007
[133] G K Scott M D Mattie C E Berger S C Benz and C CBenz ldquoRapid alteration of microRNA levels by histone deacety-lase inhibitionrdquo Cancer Research vol 66 no 3 pp 1277ndash12812006
[134] K Gajjar P L Martin-Hirsch and F L Martin ldquoCYP1B1 andhormone-induced cancerrdquoCancer Letters vol 324 no 1 pp 13ndash30 2012
[135] Y Tsuchiya M Nakajima S Takagi T Taniya and T YokoildquoMicroRNA regulates the expression of human cytochromeP450 1B1rdquoCancer Research vol 66 no 18 pp 9090ndash9098 2006
[136] L Jin O Wessely E Marcusson C Ivan G Calin and S KAlahari ldquoProoncogenic factors miR-23b and miR-27b are reg-ulated by Her2Neu EGF and TNF-120572 in breast cancerrdquo CancerResearch vol 73 pp 2884ndash2896 2013
[137] J T Buijs P Juarez and T A Guise ldquoTherapeutic strategiesto target TGF-120573 in the treatment of bone metastasesrdquo CurrentPharmaceutical Biotechnology vol 12 pp 2121ndash2137 2011
[138] T Kim and A Reitmair ldquoNon-coding RNAs functional aspectsand diagnostic utility in oncologyrdquo International Journal ofMolecular Sciences vol 14 pp 4934ndash4968 2013
[139] Y X Fang andWQGao ldquoRoles ofmicroRNAs during prostatictumorigenesis and tumor progressionrdquo Oncogene 2013
[140] J Tang A Ahmad and F H Sarkar ldquoThe role of microRNAs inbreast cancer migration invasion andmetastasisrdquo InternationalJournal of Molecular Sciences vol 13 pp 13414ndash13437 2012
[141] J B Patel H N Appaiah R M Burnett et al ldquoControl of EVI-1oncogene expression in metastatic breast cancer cells throughmicroRNA miR-22rdquo Oncogene vol 30 no 11 pp 1290ndash13012011
[142] S Valastyan A Chang N Benaich F Reinhardt and R AWeinberg ldquoActivation ofmiR-31 function in already-establishedmetastases elicits metastatic regressionrdquo Genes and Develop-ment vol 25 no 6 pp 646ndash659 2011
[143] S F Tavazoie C Alarcon T Oskarsson et al ldquoEndogenoushuman microRNAs that suppress breast cancer metastasisrdquoNature vol 451 no 7175 pp 147ndash152 2008
[144] P A Gregory A G Bert E L Paterson et al ldquoThe miR-200 family and miR-205 regulate epithelial to mesenchymaltransition by targeting ZEB1 and SIP1rdquo Nature Cell Biology vol10 no 5 pp 593ndash601 2008
[145] P Gandellini M Folini N Longoni et al ldquoMiR-205 exertstumor-suppressive functions in human prostate through down-regulation of protein kinase C120576rdquo Cancer Research vol 69 no 6pp 2287ndash2295 2009
[146] S S Myatt J Wang L J Monteiro et al ldquoDefinition ofmicroRNAs that repress expression of the tumor suppressorgene FOXO1 in endometrial cancerrdquo Cancer Research vol 70no 1 pp 367ndash377 2010
[147] QWang D-C Li Z-F Li et al ldquoUpregulation of miR-27a con-tributes to the malignant transformation of human bronchialepithelial cells induced by SV40 small T antigenrdquoOncogene vol30 no 36 pp 3875ndash3886 2011
[148] Y Wang R Rathinam A Walch and S K Alahari ldquoST14(suppression of tumorigenicity 14) gene is a target for miR-27band the inhibitory effect of ST14 on cell growth isindependentofmiR-27b regulationrdquo Journal of Biological Chemistry vol 284no 34 pp 23094ndash23106 2009
[149] N Akhtar Z Rasheed S Ramamurthy A N Anbazhagan F RVoss andTMHaqqi ldquoMicroRNA-27b regulates the expressionof matrix metalloproteinase 13 in human osteoarthritis chon-drocytesrdquoArthritis andRheumatism vol 62 no 5 pp 1361ndash13712010
[150] W Tang J Zhu S Su et al ldquoMiR-27 as a prognostic marker forbreast cancer progression and patient survivalrdquo PLoS ONE vol7 Article ID e51702 2012
[151] G Grelier N Voirin A-S Ay et al ldquoPrognostic value of Dicerexpression in human breast cancers and association with themesenchymal phenotyperdquo British Journal of Cancer vol 101 no4 pp 673ndash683 2009
[152] S Liu R H Goldstein E M Scepansky and M RosenblattldquoInhibition of Rho-associated kinase signaling prevents breastcancer metastasis to human bonerdquo Cancer Research vol 69 no22 pp 8742ndash8751 2009
[153] S Dangi-Garimella J Yun E M Eves et al ldquoRaf kinase inhib-itory protein suppresses a metastasis signalling cascade involv-ing LIN28 and let-7rdquo EMBO Journal vol 28 no 4 pp 347ndash3582009
[154] C Wang Y Tai M P Lisanti and D J Liao ldquoc-Myc inductionof programmed cell death may contribute to carcinogenesis aperspective inspired by several concepts of chemical carcino-genesisrdquo Cancer Biology andTherapy vol 11 no 7 pp 615ndash6262011
[155] Y Dorsett K M McBride M Jankovic et al ldquoMicroRNA-155suppresses activation-induced cytidine deaminase-mediatedMyc-Igh translocationrdquo Immunity vol 28 no 5 pp 630ndash6382008
[156] A Lal F Navarro C A Maher et al ldquomiR-24 inhibits cell pro-liferation by targeting E2F2MYC and other cell-cycle genes viabinding to ldquoseedlessrdquo 31015840UTRMicroRNA recognition elementsrdquoMolecular Cell vol 35 no 5 pp 610ndash625 2009
[157] L Ma J Young H Prabhala et al ldquoMiR-9 a MYCMYCN-activated microRNA regulates E-cadherin and cancer metas-tasisrdquo Nature Cell Biology vol 12 no 3 pp 247ndash256 2010
[158] P Gao I Tchernyshyov T-C Chang et al ldquoC-Myc suppressionof miR-23ab enhances mitochondrial glutaminase expressionand glutamine metabolismrdquoNature vol 458 no 7239 pp 762ndash765 2009
[159] M Sachdeva S Zhu F Wu et al ldquop53 represses c-Myc throughinduction of the tumor suppressor miR-145rdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 106 no 9 pp 3207ndash3212 2009
[160] A Arnold and A Papanikolaou ldquoCyclin D1 in breast cancerpathogenesisrdquo Journal of Clinical Oncology vol 23 no 18 pp4215ndash4224 2005
[161] D J Liao andR BDickson ldquoc-Myc in breast cancerrdquoEndocrine-Related Cancer vol 7 no 3 pp 143ndash164 2000
[162] J Chen H E Feilotter G C Pare et al ldquoMicroRNA-193brepresses cell proliferation and regulates cyclin D1 in mela-nomardquo American Journal of Pathology vol 176 no 5 pp 2520ndash2529 2010
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
Disease Markers 387
[163] W Xia J Li L Chen et al ldquoMicroRNA-200b regulates cyclinD1 expression and promotes S-phase entry by targeting RND3inHeLa cellsrdquoMolecular and Cellular Biochemistry vol 344 no1-2 pp 261ndash266 2010
[164] P Bhat-Nakshatri G Wang N R Collins et al ldquoEstradiol-regulated microRNAs control estradiol response in breast can-cer cellsrdquo Nucleic Acids Research vol 37 no 14 pp 4850ndash48612009
[165] Q Jiang M-G Feng and Y-Y Mo ldquoSystematic validation ofpredictedmicroRNAs for cyclin D1rdquo BMCCancer vol 9 article194 2009
[166] Y Onodera Y Miki T Suzuki et al ldquoRunx2 in human breastcarcinoma its potential roles in cancer progressionrdquo CancerScience vol 101 no 12 pp 2670ndash2675 2010
[167] D T Leong J Lim X Goh et al ldquoCancer-related ectopicexpression of the bone-related transcription factor RUNX2 innon-osseousmetastatic tumor cells is linked to cell proliferationand motilityrdquo Breast Cancer Research vol 12 no 5 article R892010
[168] C K Inman and P Shore ldquoThe osteoblast transcription factorRunx2 is expressed in mammary epithelial cells and mediatesosteopontin expressionrdquo Journal of Biological Chemistry vol278 no 49 pp 48684ndash48689 2003
[169] J Pratap K M Imbalzano J M Underwood et al ldquoEctopicRunx2 expression in mammary epithelial cells disrupts forma-tion of normal acini structure implications for breast cancerprogressionrdquo Cancer Research vol 69 no 17 pp 6807ndash68142009
[170] K Das D T Leong A Gupta et al ldquoPositive associationbetween nuclear Runx2 and oestrogen-progesterone receptorgene expression characterises a biological subtype of breastcancerrdquo European Journal of Cancer vol 45 no 13 pp 2239ndash2248 2009
[171] A Javed G L Barnes J Pratap et al ldquoImpaired intranucleartrafficking of Runx2 (AML3CBFA1) transcription factors inbreast cancer cells inhibits osteolysis in vivordquo Proceedings of theNational Academy of Sciences of the United States of Americavol 102 no 5 pp 1454ndash1459 2005
[172] M Tome P Lopez-Romero C Albo et al ldquoMiR-335 orches-trates cell proliferation migration and differentiation in humanmesenchymal stem cellsrdquo Cell Death and Differentiation vol 18no 6 pp 985ndash995 2011
[173] M Q Hassan J A R Gordon M M Beloti et al ldquoA networkconnecting Runx2 SATB2 and the miR-23asim27asim24-2 clusterregulates the osteoblast differentiation programrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 107 no 46 pp 19879ndash19884 2010
[174] J Huang L Zhao L Xing and D Chen ldquoMicroRNA-204 reg-ulates Runx2 protein expression and mesenchymal progenitorcell differentiationrdquo Stem Cells vol 28 no 2 pp 357ndash364 2010
[175] Z Li M Q Hassan S Volinia et al ldquoA microRNA signaturefor a BMP2-induced osteoblast lineage commitment programrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 105 no 37 pp 13906ndash13911 2008
[176] R Spizzo M S Nicoloso L Lupini et al ldquoMiR-145 participateswith TP53 in a death-promoting regulatory loop and targetsestrogen receptor-120572 in human breast cancer cellsrdquo Cell Deathand Differentiation vol 17 no 2 pp 246ndash254 2010
[177] D Barh S Parida B P Parida and G Viswanathan ldquoLet-7miR-125 miR-205 and miR-296 are prospective therapeuticagents in breast cancer molecular medicinerdquo GeneTherapy andMolecular Biology vol 12 no 2 pp 189ndash206 2008
[178] J Pratap M Galindo S K Zaidi et al ldquoCell growth regulatoryrole of Runx2 during proliferative expansion of preosteoblastsrdquoCancer Research vol 63 no 17 pp 5357ndash5362 2003
[179] J Pratap J B Lian A Javed et al ldquoRegulatory roles of Runx2in metastatic tumor and cancer cell interactions with bonerdquoCancer andMetastasis Reviews vol 25 no 4 pp 589ndash600 2006
[180] K Blyth F Vaillant L Hanlon et al ldquoRunx2 and MYC collab-orate in lymphoma development by suppressing apoptotic andgrowth arrest pathways in vivordquo Cancer Research vol 66 no 4pp 2195ndash2201 2006
[181] M Stewart A Terry M Hu et al ldquoProviral insertions inducethe expression of bone-specific isoforms of PEBP2120572A (CBFA1)evidence for a new myc collaborating oncogenerdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 94 no 16 pp 8646ndash8651 1997
[182] J Pratap J J Wixted T Gaur et al ldquoRunx2 transcriptional acti-vation of Indian Hedgehog and a downstream bone metastaticpathway in breast cancer cellsrdquo Cancer Research vol 68 no 19pp 7795ndash7802 2008
[183] R Hu W Liu H Li et al ldquoA Runx2miR-3960miR-2861 reg-ulatory feedback loop during mouse osteoblast differentiationrdquoJournal of Biological Chemistry vol 286 no 14 pp 12328ndash123392011
![Computer Architecture - Weebly...1 [RISC AND CISC] RISC AND CISC Generali 1. The dominant architecture in the PC market, the Intel IA-32, belongs to the Complex Instruction Set Computer](https://static.fdocuments.us/doc/165x107/60d6345cb801a56cc5222cad/computer-architecture-weebly-1-risc-and-cisc-risc-and-cisc-generali-1-the.jpg)
