ReseachArticle - Hindawi Publishing CorporationCYPC, CYPE, CYP A, and CYPA were all dierentially...

Research ArticleDownregulation of CYP2A6 and CYP2C8 in TumorTissues Is Linked to Worse Overall Survival and Recurrence-FreeSurvival from Hepatocellular Carcinoma

Xiaojing Ren Yuanyuan Ji Xuhua Jiang and Xun Qi

Department of Liver Disease Shanghai Public Health Clinical Center Fudan University Shanghai 201508 China

Correspondence should be addressed to Xun Qi qixunshaphcorg

Received 5 June 2018 Accepted 8 July 2018 Published 25 July 2018

Academic Editor Fumio Imazeki

Copyright copy 2018 Xiaojing Ren 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

Objective This study aimed to evaluate the links between CYP450 family genes in tumor tissues and hepatocellular carcinoma(HCC) outcomes Methods Gene Expression Omnibus (GEO) databases GSE14520 and GSE36376 were used to identifydifferential expressed CYP450 genes between tumor and nontumor tissues and related to HCC clinicopathological features andsurvivals Results Seven CYP450 genes including CYP1A2 CYP2A6 CYP2C8 CYP2C9 CYP2E1 CYP3A4 and CYP4A11 weredownregulated in tumor tissues which were validated in both GSE14520 and GSE36376 HCC patients with CYP2A6 and CYP2C8low levels in tumor tissues suffered from poorer overall survival (OS) compared to those with high CYP2A6 and CYP2C8 inGSE14520 profile (log ranks P = 001 and P = 0006 respectively) In addition HCC patients with lower CYP2A6 and CYP2C8 intumors had worse recurrence-free survival (RFS) than those with higher CYP2A6 and CYP2C8 (log ranks P = 002 and P = 0012respectively) In GSE36376 validation dataset HCC patients with lower CYP2A6 and CYP2C8 had worse OS and RFS than thosewith higher CYP2A6 andCYP2C8 (all P lt 005) in line with results in GSE14520 dataset Additionally lower CYP2A6 and CYP2C8are associated with advanced clinicopathological features including tumor staging vascular invasion intrahepatic metastasis andhigh alpha fetoprotein (all P lt 005)Conclusion Downregulation of CYP2A6 and CYP2C8 in tumor tissues links to poorer OS andRFS in HCC patients

1 Introduction

Accounting for 85-90 of primary liver malignancy hepato-cellular carcinoma (HCC) is the fifth most common tumorworldwide and the second most common cause of cancer-related death [1ndash4] Over the last 20 years the incidenceof HCC has been rising in the United States and China[1 2] Given the complexity of the disease and the largenumber of useful therapies patients with HCC still sufferedfrom poor prognosis [5] To find novel biomarkers forpredicting outcomes and efficacy of anticancer agents is ofgreat essential

The efficacy of anticancer therapy is limited by our inabil-ity to predict patient outcomes such as tumor response andtoxicity Drug-metabolizing enzymes in tumor tissues playcritical roles in the activation or inactivation of numerous

cytotoxic drugs and influence the susceptibility of host andneoplasm to their effects [6] Cytochrome P450s (CYPs) is alarge group of enzymes that localize to mitochondrial mem-branes or the endoplasmic reticulum and play crucial rolesin the metabolism of endogenous and exogenous moleculesincluding most drugs [7] It has been suggested that thelocal expression of CYPs in tumors is very important forthe management of cancer since these functionally associatedenzymes might be involved both in the development of HCCand in determining the anticancer drug sensitivity of suchtumor [8 9]

Given the key roles of CYPs in tumor developmentfew data were available of CYPs in tumor tissues and HCCclinicopathological features and survivals This study aimedto evaluate differential expressed CYP genes and to presentpredict roles of CYPs in tumor tissues for HCC survivals

HindawiBioMed Research InternationalVolume 2018 Article ID 5859415 9 pageshttpsdoiorg10115520185859415

2 BioMed Research International

2 Materials and Methods

21 Patients HCC patients in this study were from cohortsin GSE14520 and GSE36376 In GSE14520 247 HCC patientswere identified Data on gene expression could not beobtained for 22 of these and a further 5 had insufficientclinical outcome data available leading to 220 patients beingincluded in the analysis Cases consisted of patients with ahistory of hepatitis B virus (HBV) infection or HBV-relatedliver cirrhosis the diagnosis of HCC was made in all cases bytwo independent pathologists who had detailed informationon clinical presentation and pathological characteristics [10]In GSE36376 240 tumor tissues containing no necrosis orhemorrhage were available from primary HCC patients whowere treated with surgical resection or liver transplantationat Samsung Medical Center Seoul Korea from July 2000to May 2006 None of the patients received preoperativechemotherapy

22 Ethics Statement In GSE14520 all liver tissue wasobtained with informed consent from patients who under-went radical resection between 2002 and 2003 at the LiverCancer Institute and Zhongshan Hospital (Fudan Univer-sity) The study was approved by the Institutional ReviewBoard of the participating institutes [10] All participantsprovided written informed consent as reported by Roessleret al [10 11] In GSE36376 informed consent was obtainedfrom each patient and the study protocol was approved bythe Institutional Review Board of Samsung Medical CenterSeoul Korea which is in agreement with reports by Lim HYet al [12] The second analysis protocol was approved by theEthics Committee of Shanghai Public Health Clinical CenterFudan University

23 Source of Data We extracted the GSE14520 andGSE36376 microarray expression profiles from GeneExpression Omnibus (GEO httpwwwncbinlmnihgovgeo) database For GSE14520 tumor sample and microarrayprocessing were reported by Roessler et al [10 11] Geneexpression levels were calculated using the matchprobespackage in the R programming environment and the log2RMA-calculated signal intensitywas reportedDetails of the ex-periment protocols and data processing are available at httpwwwncbinlmnihgovgeoqueryacccgiacc=GSM362949For GSE36376 tumor tissues of HCC patients after curativehepatectomy were profiled using Illumina HumanHT-12V40 expression beadchip (Illumina Inc San Diego CA)The expression data was retrieved from Gene ExpressionOmnibus (GSE14520 and GSE36376 httpwwwncbinlmnihgovgeo) [10ndash12] We used GEO2R tool for identifyingdifferential expressed genes with criteria of |log FC| gt 2 andadjust P lt 00001

24 Definitions of Outcomes Overall survival (OS) wasdefined as the time from surgery to death from any dis-ease Recurrence-free survival (RFS) was defined as timefrom surgery to the date of tumor recurrence or death InGSE14520 RFS was diagnosed by new lesions found in thecomputed tomography (CT) or magnetic resonance imaging

(MRI) scans and an abnormal alpha-fetoprotein (AFP) valuewith a cut-off of 300 ngml including instanceswhen the highpretreatment AFP value did not decrease to a normal level orincreased again after becoming normal In GSE36376 patientserum AFP levels were evaluated and three-phase dynamicCT scans were performed at least once every 3 months aftersurgery until December 31 2010 When tumor recurrencewas suspected precise diagnostic imaging was performed byMRI

25 Statistical Analysis Studentrsquos t-test was used to comparemeans for normally distributed continuous data and theMannndashWhitney U test was used for nonnormally distributedcontinuous data and the Chi-squared test was used forcategorical variables The KaplanndashMeier method was usedto compare survivals between different groups and the log-rank test was used to estimate the difference in survivalStatistical analyses were performed using PASW Statisticssoftware version 230 from SPSS Inc (Chicago IL USA) Atwo-tailed P lt 005 was considered statistically significant

3 Results

31 Differential Expressed CYPs in Both GSE14520 andGSE36376 As shown in Table 1 eleven CYPs includingCYP1A2 CYP2A6 CYP2B6 CYP2B7P CYP2C8 CYP2C9CYP2E1 CYP3A4 CYP4A11 CYP4A22 and CYP39A1were identified differential expression in GSE14520 data-set In GSE36376 eight CYPs including CYP1A2 CYP2A6CYP2C8 CYP2C9 CYP2E1 CYP3A4 CYP4A11 andCYP8B1 were significantly differentially expressed Com-binedly seven CYPs including CYP1A2 CYP2A6 CYP2C8CYP2C9 CYP2E1 CYP3A4 and CYP4A11 were alldifferentially expressed in both GSE14520 and GSE36376datasets As shown in Figures 1 and 2 all the seven differentialCYPs were lower expressed in tumor tissues compared tothose in nontumor tissues

32 CYP Genes Associated with OS in HCC In GSE14520dataset we identified the fact that low expressions offour CYPs including CYP2A6 CYP2C8 CYP2E1 andCYP4A11 in tumor tissues were significantly associated withworse OS in HCC patients (log ranks P = 001 00060024 and 0007 respectively Figure 3) Furtherly we con-ducted KaplanndashMeier curve for validation of the four CYPs(CYP2A6 CYP2C8 CYP2E1 and CYP4A11) for evaluatingits links to OS in HCC patients from GSE36376 datasetAs shown in Figure 4 downregulation of CYP2A6 wassignificantly associated with OS in HCC patients with cut-off of 100 detected by R program (log rank P = 0015Figure 4(a)) And low expression of CYP2C8 in tumor tissueswas linked to worse OS in HCC patients with median cut-off(log rank P = 0018 Figure 4(b))

33 CYP Genes Associated with RFS in HCC In GSE14520dataset low expressions of two CYPs including CYP2A6and CYP2C8 were significantly associated with worse RFSin HCC patients (log ranks P = 002 and 0012 respectively

BioMed Research International 3

Table1Differentia

lexpressed

CYP4

50genesb

etwe

entumor

andno

ntum

ortissues

from

GSE

14520andGSE

36376

Genes

tB

logF

CPvalue

AdjustPvalue

Com

mon

genes

GSE

14520

CYP1A2

CYP2

A6

CYP2

C8CY

P2C9

CYP2

E1CY

P3A4

CYP4

A11

CYP1A2

257352762

49901907

444

705742

163E

-22

731E-27

CYP2

A6

79627779

12348032

307188278

366

E-08

766E

-10

CYP2

B6157936233

33005014

389306938

665E-16

537E-19

CYP2

B7P

24284837

180032064

35900

407

18E-81

125E

-83

CYP2

C864917335

7665054

432698325

000

00018

878E-08

CYP2

C92337

15192

46568366

336577273

163E

-21

293E-25

CYP2

E182816633

13340

866

473015789

000

0000

016

281E-10

CYP3

A4

799706

641245527

399461005

337E-08

687E-10

CYP4

A11

88192119

14989899

311363397

399E-09

529E-11

CYP4

A22

864

45676

14457718

317968421

61E-09

906E

-11

CYP3

9A1

105703094

20114355

319955263

584E-11

294E-13

GSE

36376

CYP1A2

30500

6424

24040

8848

497

129E

-106

605E-110

CYP2

A6

150803249

83643956

247

271E-40

13E-41

CYP2

C817410711

107141675

305

25E

-50

736E

-52

CYP2

C9149361339

82220342

236

11E-39

544

E-41

CYP2

E1150716592

83558278

3295E-40

142E

-41

CYP3

A4

2247200

681595

80681

391

13E-72

103E

-74

CYP4

A11

143069129

76061125

206

462E-37

265E-38

CYP8

B1129803487

63406

73218

118E

-31

89E

-33


Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C9

Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C8

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP2

A6

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP1

A2

Adjacent Tumor

Plt00001

minus1000minus500

050010001500200025003000

CYP4

A11

Adjacent Tumor

Plt00001

minus2000

0

2000

4000

6000

8000

10000

CYP3

A4

Adjacent Tumor

Plt00001

minus3000

0

3000

6000

9000

12000

15000

CYP2

E1

Figure 1 Differential expressed CYPs in GSE14520 profile

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

0246810121416

CYP1

A2

02468101214161820

CYP2

E1

0246810121416

CYP2

A6

0246810121416

CYP3

A4

02468101214161820

CYP2

C8

02468101214161820

CYP4

A11

0246810121416

CYP2

C9


Figure 5) In validation set from GSE36376 downregulationof CYP2A6 and CYP2C8 was also related to poorer RFS inHCCpatients (log ranks P = 0008 and P lt 0001 respectivelyFigure 6)

34 Relationship between CYP2A6 CYP2C8 and HCC Clini-copathological Features In GSE14520 profile HCC patientshad advanced TNM staging in CYP2A6 and CYP2C8 lowexpression groups compared to those in CYP2A6 andCYP2C8 high expression groups (P = 0005 and P = 0032respectively Table 2) Interestingly in GSE36376 databaseHCCpatients with lowCYP2A6 and CYP2C8 levels in tumortissues suffered from more advanced edmondson grade andAJCC staging (all P lt 005 Table 2) Additionally HCC

patients with low CYP2A6 and CYP2C8 levels in tumortissues had higher risk of vascular invasion major portalvein invasion and intrahepatic metastasis (all P lt 005Table 2) Also those HCC cases with low CYP2A6 andCYP2C8 had significantly higher AFP level (P lt 0001Table 2) Considered above we assumed that downregulationof CYP2A6 and CYP2C8 in tumor tissues is associated withworse clinicopathological characteristics in HCC patients

4 Discussion

As key enzymes in cancer formation and cancer treatmentCYPs mediate the metabolic activation of multiple procar-cinogens and participate in the inactivation and activation


Table2Clinicop

atho

logicalcom

paris

ongrou

pedby

CYP2

A6andCY

P2C8

from

GSE

14520andGSE

36376

Category

CYP2

A6

Pvalue

CYP2

C8Pvalue

High

Low

High

Low

GSE

14520

HBV

viralstatus(AV

R-CC

CC

NA)

25814

31745

0635

28784

28775

0943

Alanine

aminotransferase

(gt50lt50U

L)

3872

5159

0099

4268

4763

0583

Maintumor

size(

gt5lt5NAcm)

34760

44651

016

33770

45641

0091

Multin

odular

(yesno)

1892

2783

0181

2189

2486

0738

Cirrho

sis(yesno)

1019

1019

101019

1019

10TN

Msta

ging

(III

IIIN

A)

5737151

3640331

000

55633210

3744272

0032

BCLC

staging

(0-A

BC

NA)

83971

7513

211

0039

79813

07914

152

0357

GSE

36376

Edmon

dson

grade(

IIII

II)1610

04

89715

001

19965

510114

000

2Tu

mor

size(

median)m

m34

(31)

40(45)

0203

34(30)

42(45)

0081

Vascular

invasio

n(yesno)

5169

8436

lt000

15169

8436

lt000

1Major

portalvein

invasio

n(yesno)

1119

8112

003

611

198112

003

6Intrahepaticmetastasis

(yesno)

1710

33882

000

21810

23783

000

5Multic

entricoccurrence

(yesno)

8112

5115

057

7113

6114

10Dire

ctinvasio

nof

adjacent

organ(yesno)

3117

2118

103117

2118

10AJC

Csta

ging

(1234)

664383

3657252

lt000

1674283

3558252

lt000

1BC

LCsta

ging

(ABC

)75432

64488

0092

80382

59538

000

9Alpha

fetoprotein[M

edian(IQ

R)ngml]

965(822)

2566(41368)

lt000

191

(616

)3038(953445)

lt000

1NAn

otavailableAV

R-CC

activev

iralreplicationchronicc

arrie

rCC

chron

iccarrierIQ

Rinterquartile

range


Mean survival monthsLow CYP2A6 group = 449plusmn24High CYP2A6 group = 532plusmn22Log rank P = 001

Low CYP2A6 (n=110)High CYP2A6 (n=110)

20 40 600Overall survival months

0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)

Mean survival monthsLow CYP2C8 group = 440plusmn25High CYP2C8 group = 541plusmn21Log rank P = 0006

Low CYP2C8 (n=110)High CYP2C8 (n=110)

0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)

Mean survival monthsLow CYP2E1 group = 450plusmn25High CYP2E1 group = 532plusmn21Log rank P = 0024

Low CYP2E1 (n=110)High CYP2E1 (n=110)


0

20

40

60

80

100

Ove

rall

surv

ival

()

(c)




0

20

40

60

80

100O

vera

ll su

rviv

al (

)

(d)

Figure 3 Overall survival (OS) comparison grouped by CYP2A6 (a) CYP2C8 (b) CYP2E1 (c) and CYP4A11 (d) from GSE14520

of anticancer drugs [13] CYPs polymorphisms have beenextensively studied with respect to genetic predispositionto cancer and clinical outcome in terms of response andtoxicity to anticancer drugs [14] Various studies have alsoshown the significance of CYPs polymorphisms in cancersusceptibility [15ndash18] Some members of CYPs family alsoaffect the metabolism of various environmental carcinogens[14] In HCC patients CYP1A1 CYP2D6 and CYP2E1 wereall found to be associatedwith increasedHCC risk in differentpopulation [19ndash21] However a consistent view does not yetexist [14 22]

CYPs are key enzymes involved in cancer developmentand mediate the metabolic activation of numerous procar-cinogens [13] To assess the CYPs activity changes would beuseful not only for designing personalized HCC treatments

but also for identifying potential factors that contribute toHCC susceptibility [23] In our study we found that sev-eral CYPs including CYP1A2 CYP2A6 CYP2C8 CYP2C9CYP2E1 CYP3A4 and CYP4A11 were all downregulatedin tumor tissues in HCC patients And low expression ofCYP2A6 in tumor tissues contributed to worse survivalsfrom HCC cases CYP2A6 is primarily expressed in the liverand metabolizes several clinically relevant substrates [24]Variation in CYP2A6 activity is involved in the metabolismor bioactivation of clinical therapeutics and carcinogensleading to an important clinical consideration [25ndash27]CYP2A6 was also associated with worse clinicopathologicalcharacteristics including advanced tumor staging vascularinvasion major portal vein invasion intrahepatic metastasisand increased AFP level in our study In line with our




20 40 60 80 100 1200Overall survival months

0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)

Figure 4 Validation of CYP2A6 (a) and CYP2C8 (b) for OS in database GSE36376



20 40 600Recurrence-free survival months

0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)

Figure 5 Recurrence-free survival (RFS) comparison grouped by CYP2A6 (a) and CYP2C8 (b)

findings CYP2A6 is highly polymorphic and its geneticvariants can result in reduced expression by affecting tran-scriptional or translational processes [28] Clinically geneticvariation in CYP2A6 may contribute to lung cancer risk[29 30] However few studies focused on the relationshipsbetween CYP2A6 and HCC development Raunio H et alinvestigated the distribution of the CYP2A6 protein in aseries of 24 human hepatocellular carcinoma (HCC) samplesby immunohistochemical analysis They found that CYP2A6protein was very heterogeneous in tumor cells and HCC donot uniformly overexpress the CYP2A6 protein suggesting

that increased expression of CYP2A6 occurred in a distinctsubpopulation of neoplastic cells Additionally they foundthat patients with CYP2A6-positive tumors achieved a morefavorable prognosis compared with patients with CYP2A6-negative tumors [31] Our results conformed and replenishedthe previous research [8 31] More detailed studies on theassociation betweenCYP2A6 andHCC should be conducted

Several genetic polymorphisms inCYP2C8may influencesurvival after cancer diagnosis due to their role in themetabolism of various breast cancer chemotherapy drugs[32] However the risk of developing colorectal cancer does




20 40 60 80 100 1200Recurrence-free survival months

0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)


Mean survival monthsLow CYP2C8 group = 406plusmn43High CYP2C8 group = 631plusmn48Log rank P lt 00010

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

12040 60 80 100200Recurrence-free survival months

(b)

Figure 6 Validation of CYP2A6 (a) and CYP2C8 (b) for RFS in database GSE36376

not seem to be related to the commonest functional geneticvariation in the CYP2C8 gene [33] Unfortunately knowl-edge on CYP2C8 in tumors and HCC outcomes is limitedOur research demonstrated that CYP2C8 downregulation intumor tissueswas associatedwith advanced tumor phenotypeand worse OS and RFS Yan et al determined seven CYPsincluding CYP2C8 in tumor and pericarcinomatous tissuesharvested from 26 patients with HBV-positive HCC usingprobe substrates A major decrease of CYP2C8 in tumortissues was observed [6] in line with a study by Zhou etal [23] Experimentally polycyclic aromatic hydrocarbons3-methylcholanthrene suppressed CYP2C8 mRNA levels inthe HCC cell line HepG2 and basal CYP2C8 expressionwas extremely low [34] That is CYP2C8 expression levelsare greatly disrupted by the tumorigenic process [6] Sincedrug metabolism mediated by tumor CYPs can be used asa marker for potential mechanism of drug resistance andoran approach to achieve optimal chemotherapy to upregulateCYP2C8 levels and activations should be considered forHCC treatment strategy

There are some limitations in this study Firstly CYPsexpression in our research is limited in mRNA level whichmay not necessarily reflect that in activities of CYPs Sec-ondly in our analysis based on GEO database no furthermechanism data were shown

According to our results we cautiously drew the conclu-sion that downregulation of CYP2A6 and CYP2C8 in tumortissues is linked to worse overall survival and recurrence-free survival from hepatocellular carcinoma A detailedunderstanding of the differential expression and activity ofCYPs within HCC tumor tissues may provide opportunitiesand more alternatives for improved therapeutic outcome

Data Availability

All the data in this study were available from GEO database(httpswwwncbinlmnihgovgeo)

Disclosure

Thisworkwas not supported by any pharmaceutical companyor government agency or grants from other sources

Conflicts of Interest

The authors declare that they have no conflicts of interest

References

[1] J K Heimbach L M Kulik R S Finn et al ldquoAASLDguidelinesfor the treatment of hepatocellular carcinomardquoHepatology vol67 no 1 pp 358ndash380 2018

[2] M Omata A-L Cheng N Kokudo et al ldquoAsiandashPacific clinicalpractice guidelines on the management of hepatocellular carci-noma a 2017 updaterdquoHepatology International vol 11 no 4 pp317ndash370 2017

[3] R L Siegel K D Miller and A Jemal ldquoCancer statistics 2017rdquoCA A Cancer Journal for Clinicians vol 67 no 1 pp 7ndash30 2017

[4] L A Torre F Bray R L Siegel J Ferlay and J Lortet-Tieulent ldquoGlobal cancer statistics 2012rdquo CA A Cancer Journalfor Clinicians vol 65 no 2 pp 87ndash108 2015

[5] P Fitzmorris M Shoreibah B S Anand and A K SingalldquoManagement of hepatocellular carcinomardquo Journal of CancerResearch and Clinical Oncology vol 141 no 5 pp 861ndash876 2015

[6] T Yan L Lu C Xie et al ldquoSeverely impaired and dysregulatedcytochrome P450 expression and activities in hepatocellularcarcinoma Implications for personalized treatment in patientsrdquo


Molecular Cancer Therapeutics vol 14 no 12 pp 2874ndash28862015

[7] S A Sheweita ldquoDrug-metabolizing enzymes mechanisms andfunctionsrdquo Current Drug Metabolism vol 1 no 2 pp 107ndash1322000

[8] T Oyama N Kagawa N Kunugita et al ldquoExpression ofcytochrome P450 in tumor tissues and its association withcancer developmentrdquo Frontiers in Bioscience vol 9 pp 1967ndash1976 2004

[9] MMichael andMM Doherty ldquoDrugmetabolism by tumoursIts nature relevance and therapeutic implicationsrdquo ExpertOpinion onDrugMetabolismampToxicology vol 3 no 6 pp 783ndash803 2007

[10] S Roessler H-L Jia A Budhu et al ldquoA unique metastasisgene signature enables prediction of tumor relapse in early-stage hepatocellular carcinoma patientsrdquo Cancer Research vol70 no 24 pp 10202ndash10212 2010

[11] S Roessler E L Long A Budhu et al ldquoIntegrative genomicidentification of genes on 8p associated with hepatocellularcarcinoma progression and patient survivalrdquo Gastroenterologyvol 142 no 4 pp 957 e912ndash966 e912 2012

[12] H-Y Lim I Sohn S Deng et al ldquoPrediction of disease-free survival in hepatocellular carcinoma by gene expressionprofilingrdquo Annals of Surgical Oncology vol 20 no 12 pp 3747ndash3753 2013

[13] C Rodriguez-Antona and M Ingelman-Sundberg ldquoCyto-chrome P450 pharmacogenetics and cancerrdquoOncogene vol 25no 11 pp 1679ndash1691 2006

[14] B Mittal S Tulsyan S Kumar R D Mittal and G AgarwalldquoCytochrome P450 in Cancer Susceptibility and TreatmentrdquoAdvances in Clinical Chemistry vol 71 pp 77ndash139 2015

[15] L Yao H C Wang J Z Liu and Z M Xiong ldquoQuantitativeassessment of the influence of cytochrome P450 2C19 genepolymorphisms and digestive tract cancer riskrdquo Tumor Biologyvol 34 no 5 pp 3083ndash3091 2013

[16] F-F Shen F-Y Zhou Q-S Xue et al ldquoAssociation betweenCYP1A1 polymorphisms and esophageal cancer A meta-Analysisrdquo Molecular Biology Reports vol 40 no 10 pp 6035ndash6042 2013

[17] K P Economopoulos and T N Sergentanis ldquoThree polymor-phisms in cytochrome P450 1B1 (CYP1B1) gene and breastcancer risk A meta-analysisrdquo Breast Cancer Research andTreatment vol 122 no 2 pp 545ndash551 2010

[18] A Abbas K Delvinquiere M Lechevrel P Lebailly P Gaudu-chon G Launoy et al ldquoGSTM1 GSTT1 GSTP1 and CYP1A1genetic polymorphisms and susceptibility to esophageal cancerin a french population different pattern of squamous cell carci-noma and adenocarcinomardquoWorld Journal of Gastroenterologyvol 10 no 23 pp 3389ndash3393 2004

[19] J A G Agundez M C Leedesma J Benıtez et al ldquoCYP2D6genes and risk of liver cancerrdquoThe Lancet vol 345 no 8953 pp830-831 1995

[20] M-W Yu A Gladek-Yarborough and S ChiamprasertldquoCytochrome P450 2E1 and glutathione S-transferase M1 poly-morphisms and susceptibility to hepatocellular carcinomardquoGastroenterology vol 109 no 4 pp 1266ndash1273 1995

[21] M-W Yu Y-H Chiu S-Y Yang et al ldquoCytochrome P450 1A1genetic polymorphisms and risk of hepatocellular carcinomaamong chronic hepatitis B carriersrdquo British Journal of Cancervol 80 no 3-4 pp 598ndash603 1999

[22] L Silvestri L Sonzogni A De Silvestri et al ldquoCYP enzymepolymorphisms and susceptibility to HCV-related chronic liverdisease and liver cancerrdquo International Journal of Cancer vol104 no 3 pp 310ndash317 2003

[23] J ZhouQWen S-F Li et al ldquoSignificant change of cytochromeP450s activities in patients with hepatocellular carcinomardquoOncotarget vol 7 no 31 pp 50612ndash50623 2016

[24] E M McDonagh C Wassenaar S P David et al ldquoPhar-mGKB summary Very important pharmacogene informationfor cytochrome P-450 family 2 subfamily A polypeptide 6rdquoPharmacogenetics and Genomics vol 22 no 9 pp 695ndash7082012

[25] K Ikeda K Yoshisue E Matsushima et al ldquoBioactivation oftegafur to 5-fluorouracil is catalyzed by cytochrome P-450 2A6in human liver microsomes in vitrordquo Clinical Cancer Researchvol 6 no 11 pp 4409ndash4415 2000

[26] H L Wong S E Murphy and S S Hecht ldquoCytochromeP450 2A-catalyzed metabolic activation of structurally simi-lar carcinogenic nitrosamines N1015840-Nitrosonornicotine enanti-omers N-nitrosopiperidine and N-nitrosopyrrolidinerdquo Chem-ical Research in Toxicology vol 18 no 1 pp 61ndash69 2005

[27] K Murai H Yamazaki K Nakagawa R Kawai and TKamataki ldquoDeactivation of anti-cancer drug letrozole to acarbinol metabolite by polymorphic cytochrome P450 2A6 inhuman liver microsomesrdquo Xenobiotica vol 39 no 11 pp 795ndash802 2009

[28] Y M Di V D-W Chow L-P Yang and S-F Zhou ldquoStructurefunction regulation and polymorphism of human cytochromeP450 2A6rdquoCurrent DrugMetabolism vol 10 no 7 pp 754ndash7802009

[29] J-M YuanHHNelson S G Carmella et al ldquoCYP2A6 geneticpolymorphisms and biomarkers of tobacco smoke constituentsin relation to risk of lung cancer in the Singapore ChineseHealth Studyrdquo Carcinogenesis vol 38 no 4 pp 411ndash418 2017

[30] Y-L Liu Y Xu F Li H Chen and S-L Guo ldquoCYP2A6deletionpolymorphism is associated with decreased susceptibility oflung cancer in Asian smokers Ameta-analysisrdquo Tumor Biologyvol 34 no 5 pp 2651ndash2657 2013

[31] H Raunio R Juvonen M Pasanen O Pelkonen P Paakkoand Y Soini ldquoCytochrome P4502A6 (CYP2A6) expression inhuman hepatocellular carcinomardquoHepatology vol 27 no 2 pp427ndash432 1998

[32] H Jernstrom E Bgeman C Rose P-E Jonsson and C IngvarldquoCYP2C8 and CYP2C9 polymorphisms in relation to tumourcharacteristics and early breast cancer related events among 652breast cancer patientsrdquo British Journal of Cancer vol 101 no 11pp 1817ndash1823 2009

[33] J M Ladero J A G Agundez C Martınez G Amo P Ayusoand E Garcıa-Martın ldquoAnalysis of the functional polymor-phism in the cytochrome P450 CYP2C8 gene rs11572080 withregard to colorectal cancer riskrdquo Frontiers in Genetics vol 3 p278 2012

[34] R Utgikar and D S Riddick ldquoDownregulation of cytochromeP450 2C8 by 3-methylcholanthrene in human hepatocellularcarcinoma cell linesrdquo Canadian Journal of Physiology andPharmacology vol 95 no 6 pp 768ndash771 2017

Stem Cells International

Hindawiwwwhindawicom Volume 2018


MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine


Behavioural Neurology

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom


2 Materials and Methods

21 Patients HCC patients in this study were from cohortsin GSE14520 and GSE36376 In GSE14520 247 HCC patientswere identified Data on gene expression could not beobtained for 22 of these and a further 5 had insufficientclinical outcome data available leading to 220 patients beingincluded in the analysis Cases consisted of patients with ahistory of hepatitis B virus (HBV) infection or HBV-relatedliver cirrhosis the diagnosis of HCC was made in all cases bytwo independent pathologists who had detailed informationon clinical presentation and pathological characteristics [10]In GSE36376 240 tumor tissues containing no necrosis orhemorrhage were available from primary HCC patients whowere treated with surgical resection or liver transplantationat Samsung Medical Center Seoul Korea from July 2000to May 2006 None of the patients received preoperativechemotherapy

22 Ethics Statement In GSE14520 all liver tissue wasobtained with informed consent from patients who under-went radical resection between 2002 and 2003 at the LiverCancer Institute and Zhongshan Hospital (Fudan Univer-sity) The study was approved by the Institutional ReviewBoard of the participating institutes [10] All participantsprovided written informed consent as reported by Roessleret al [10 11] In GSE36376 informed consent was obtainedfrom each patient and the study protocol was approved bythe Institutional Review Board of Samsung Medical CenterSeoul Korea which is in agreement with reports by Lim HYet al [12] The second analysis protocol was approved by theEthics Committee of Shanghai Public Health Clinical CenterFudan University

23 Source of Data We extracted the GSE14520 andGSE36376 microarray expression profiles from GeneExpression Omnibus (GEO httpwwwncbinlmnihgovgeo) database For GSE14520 tumor sample and microarrayprocessing were reported by Roessler et al [10 11] Geneexpression levels were calculated using the matchprobespackage in the R programming environment and the log2RMA-calculated signal intensitywas reportedDetails of the ex-periment protocols and data processing are available at httpwwwncbinlmnihgovgeoqueryacccgiacc=GSM362949For GSE36376 tumor tissues of HCC patients after curativehepatectomy were profiled using Illumina HumanHT-12V40 expression beadchip (Illumina Inc San Diego CA)The expression data was retrieved from Gene ExpressionOmnibus (GSE14520 and GSE36376 httpwwwncbinlmnihgovgeo) [10ndash12] We used GEO2R tool for identifyingdifferential expressed genes with criteria of |log FC| gt 2 andadjust P lt 00001

24 Definitions of Outcomes Overall survival (OS) wasdefined as the time from surgery to death from any dis-ease Recurrence-free survival (RFS) was defined as timefrom surgery to the date of tumor recurrence or death InGSE14520 RFS was diagnosed by new lesions found in thecomputed tomography (CT) or magnetic resonance imaging

(MRI) scans and an abnormal alpha-fetoprotein (AFP) valuewith a cut-off of 300 ngml including instanceswhen the highpretreatment AFP value did not decrease to a normal level orincreased again after becoming normal In GSE36376 patientserum AFP levels were evaluated and three-phase dynamicCT scans were performed at least once every 3 months aftersurgery until December 31 2010 When tumor recurrencewas suspected precise diagnostic imaging was performed byMRI

25 Statistical Analysis Studentrsquos t-test was used to comparemeans for normally distributed continuous data and theMannndashWhitney U test was used for nonnormally distributedcontinuous data and the Chi-squared test was used forcategorical variables The KaplanndashMeier method was usedto compare survivals between different groups and the log-rank test was used to estimate the difference in survivalStatistical analyses were performed using PASW Statisticssoftware version 230 from SPSS Inc (Chicago IL USA) Atwo-tailed P lt 005 was considered statistically significant

3 Results

31 Differential Expressed CYPs in Both GSE14520 andGSE36376 As shown in Table 1 eleven CYPs includingCYP1A2 CYP2A6 CYP2B6 CYP2B7P CYP2C8 CYP2C9CYP2E1 CYP3A4 CYP4A11 CYP4A22 and CYP39A1were identified differential expression in GSE14520 data-set In GSE36376 eight CYPs including CYP1A2 CYP2A6CYP2C8 CYP2C9 CYP2E1 CYP3A4 CYP4A11 andCYP8B1 were significantly differentially expressed Com-binedly seven CYPs including CYP1A2 CYP2A6 CYP2C8CYP2C9 CYP2E1 CYP3A4 and CYP4A11 were alldifferentially expressed in both GSE14520 and GSE36376datasets As shown in Figures 1 and 2 all the seven differentialCYPs were lower expressed in tumor tissues compared tothose in nontumor tissues

32 CYP Genes Associated with OS in HCC In GSE14520dataset we identified the fact that low expressions offour CYPs including CYP2A6 CYP2C8 CYP2E1 andCYP4A11 in tumor tissues were significantly associated withworse OS in HCC patients (log ranks P = 001 00060024 and 0007 respectively Figure 3) Furtherly we con-ducted KaplanndashMeier curve for validation of the four CYPs(CYP2A6 CYP2C8 CYP2E1 and CYP4A11) for evaluatingits links to OS in HCC patients from GSE36376 datasetAs shown in Figure 4 downregulation of CYP2A6 wassignificantly associated with OS in HCC patients with cut-off of 100 detected by R program (log rank P = 0015Figure 4(a)) And low expression of CYP2C8 in tumor tissueswas linked to worse OS in HCC patients with median cut-off(log rank P = 0018 Figure 4(b))

33 CYP Genes Associated with RFS in HCC In GSE14520dataset low expressions of two CYPs including CYP2A6and CYP2C8 were significantly associated with worse RFSin HCC patients (log ranks P = 002 and 0012 respectively


Table1Differentia

lexpressed

CYP4

50genesb

etwe

entumor

andno

ntum

ortissues

from

GSE

14520andGSE

36376

Genes

tB

logF

CPvalue

AdjustPvalue

Com

mon

genes

GSE

14520

CYP1A2

CYP2

A6

CYP2

C8CY

P2C9

CYP2

E1CY

P3A4

CYP4

A11

CYP1A2

257352762

49901907

444

705742

163E

-22

731E-27

CYP2

A6

79627779

12348032

307188278

366

E-08

766E

-10

CYP2

B6157936233

33005014

389306938

665E-16

537E-19

CYP2

B7P

24284837

180032064

35900

407

18E-81

125E

-83

CYP2

C864917335

7665054

432698325

000

00018

878E-08

CYP2

C92337

15192

46568366

336577273

163E

-21

293E-25

CYP2

E182816633

13340

866

473015789

000

0000

016

281E-10

CYP3

A4

799706

641245527

399461005

337E-08

687E-10

CYP4

A11

88192119

14989899

311363397

399E-09

529E-11

CYP4

A22

864

45676

14457718

317968421

61E-09

906E

-11

CYP3

9A1

105703094

20114355

319955263

584E-11

294E-13

GSE

36376

CYP1A2

30500

6424

24040

8848

497

129E

-106

605E-110

CYP2

A6

150803249

83643956

247

271E-40

13E-41

CYP2

C817410711

107141675

305

25E

-50

736E

-52

CYP2

C9149361339

82220342

236

11E-39

544

E-41

CYP2

E1150716592

83558278

3295E-40

142E

-41

CYP3

A4

2247200

681595

80681

391

13E-72

103E

-74

CYP4

A11

143069129

76061125

206

462E-37

265E-38

CYP8

B1129803487

63406

73218

118E

-31

89E

-33


Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C9

Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C8

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP2

A6

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP1

A2

Adjacent Tumor

Plt00001

minus1000minus500

050010001500200025003000

CYP4

A11

Adjacent Tumor

Plt00001

minus2000

0

2000

4000

6000

8000

10000

CYP3

A4

Adjacent Tumor

Plt00001

minus3000

0

3000

6000

9000

12000

15000

CYP2

E1


Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

0246810121416

CYP1

A2

02468101214161820

CYP2

E1

0246810121416

CYP2

A6

0246810121416

CYP3

A4

02468101214161820

CYP2

C8

02468101214161820

CYP4

A11

0246810121416

CYP2

C9





4 Discussion



Table2Clinicop

atho

logicalcom

paris

ongrou

pedby

CYP2

A6andCY

P2C8

from

GSE

14520andGSE

36376

Category

CYP2

A6

Pvalue

CYP2

C8Pvalue

High

Low

High

Low

GSE

14520

HBV

viralstatus(AV

R-CC

CC

NA)

25814

31745

0635

28784

28775

0943

Alanine

aminotransferase

(gt50lt50U

L)

3872

5159

0099

4268

4763

0583

Maintumor

size(

gt5lt5NAcm)

34760

44651

016

33770

45641

0091

Multin

odular

(yesno)

1892

2783

0181

2189

2486

0738

Cirrho

sis(yesno)

1019

1019

101019

1019

10TN

Msta

ging

(III

IIIN

A)

5737151

3640331

000

55633210

3744272

0032

BCLC

staging

(0-A

BC

NA)

83971

7513

211

0039

79813

07914

152

0357

GSE

36376

Edmon

dson

grade(

IIII

II)1610

04

89715

001

19965

510114

000

2Tu

mor

size(

median)m

m34

(31)

40(45)

0203

34(30)

42(45)

0081

Vascular

invasio

n(yesno)

5169

8436

lt000

15169

8436

lt000

1Major

portalvein

invasio

n(yesno)

1119

8112

003

611

198112

003


(yesno)

1710

33882

000

21810

23783

000

5Multic

entricoccurrence

(yesno)

8112

5115

057

7113

6114

10Dire

ctinvasio

nof

adjacent

organ(yesno)

3117

2118

103117

2118

10AJC

Csta

ging

(1234)

664383

3657252

lt000

1674283

3558252

lt000

1BC

LCsta

ging

(ABC

)75432

64488

0092

80382

59538

000

9Alpha

fetoprotein[M

edian(IQ

R)ngml]

965(822)

2566(41368)

lt000

191

(616

)3038(953445)

lt000

1NAn

otavailableAV

R-CC

activev


arrie

rCC

chron

iccarrierIQ

Rinterquartile

range





0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)




0

20

40

60

80

100

Ove

rall

surv

ival

()

(c)




0

20

40

60

80

100O

vera

ll su

rviv

al (

)

(d)









0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)





0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)









0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















Table1Differentia

lexpressed

CYP4

50genesb

etwe

entumor

andno

ntum

ortissues

from

GSE

14520andGSE

36376

Genes

tB

logF

CPvalue

AdjustPvalue

Com

mon

genes

GSE

14520

CYP1A2

CYP2

A6

CYP2

C8CY

P2C9

CYP2

E1CY

P3A4

CYP4

A11

CYP1A2

257352762

49901907

444

705742

163E

-22

731E-27

CYP2

A6

79627779

12348032

307188278

366

E-08

766E

-10

CYP2

B6157936233

33005014

389306938

665E-16

537E-19

CYP2

B7P

24284837

180032064

35900

407

18E-81

125E

-83

CYP2

C864917335

7665054

432698325

000

00018

878E-08

CYP2

C92337

15192

46568366

336577273

163E

-21

293E-25

CYP2

E182816633

13340

866

473015789

000

0000

016

281E-10

CYP3

A4

799706

641245527

399461005

337E-08

687E-10

CYP4

A11

88192119

14989899

311363397

399E-09

529E-11

CYP4

A22

864

45676

14457718

317968421

61E-09

906E

-11

CYP3

9A1

105703094

20114355

319955263

584E-11

294E-13

GSE

36376

CYP1A2

30500

6424

24040

8848

497

129E

-106

605E-110

CYP2

A6

150803249

83643956

247

271E-40

13E-41

CYP2

C817410711

107141675

305

25E

-50

736E

-52

CYP2

C9149361339

82220342

236

11E-39

544

E-41

CYP2

E1150716592

83558278

3295E-40

142E

-41

CYP3

A4

2247200

681595

80681

391

13E-72

103E

-74

CYP4

A11

143069129

76061125

206

462E-37

265E-38

CYP8

B1129803487

63406

73218

118E

-31

89E

-33


Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C9

Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C8

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP2

A6

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP1

A2

Adjacent Tumor

Plt00001

minus1000minus500

050010001500200025003000

CYP4

A11

Adjacent Tumor

Plt00001

minus2000

0

2000

4000

6000

8000

10000

CYP3

A4

Adjacent Tumor

Plt00001

minus3000

0

3000

6000

9000

12000

15000

CYP2

E1


Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

0246810121416

CYP1

A2

02468101214161820

CYP2

E1

0246810121416

CYP2

A6

0246810121416

CYP3

A4

02468101214161820

CYP2

C8

02468101214161820

CYP4

A11

0246810121416

CYP2

C9





4 Discussion



Table2Clinicop

atho

logicalcom

paris

ongrou

pedby

CYP2

A6andCY

P2C8

from

GSE

14520andGSE

36376

Category

CYP2

A6

Pvalue

CYP2

C8Pvalue

High

Low

High

Low

GSE

14520

HBV

viralstatus(AV

R-CC

CC

NA)

25814

31745

0635

28784

28775

0943

Alanine

aminotransferase

(gt50lt50U

L)

3872

5159

0099

4268

4763

0583

Maintumor

size(

gt5lt5NAcm)

34760

44651

016

33770

45641

0091

Multin

odular

(yesno)

1892

2783

0181

2189

2486

0738

Cirrho

sis(yesno)

1019

1019

101019

1019

10TN

Msta

ging

(III

IIIN

A)

5737151

3640331

000

55633210

3744272

0032

BCLC

staging

(0-A

BC

NA)

83971

7513

211

0039

79813

07914

152

0357

GSE

36376

Edmon

dson

grade(

IIII

II)1610

04

89715

001

19965

510114

000

2Tu

mor

size(

median)m

m34

(31)

40(45)

0203

34(30)

42(45)

0081

Vascular

invasio

n(yesno)

5169

8436

lt000

15169

8436

lt000

1Major

portalvein

invasio

n(yesno)

1119

8112

003

611

198112

003


(yesno)

1710

33882

000

21810

23783

000

5Multic

entricoccurrence

(yesno)

8112

5115

057

7113

6114

10Dire

ctinvasio

nof

adjacent

organ(yesno)

3117

2118

103117

2118

10AJC

Csta

ging

(1234)

664383

3657252

lt000

1674283

3558252

lt000

1BC

LCsta

ging

(ABC

)75432

64488

0092

80382

59538

000

9Alpha

fetoprotein[M

edian(IQ

R)ngml]

965(822)

2566(41368)

lt000

191

(616

)3038(953445)

lt000

1NAn

otavailableAV

R-CC

activev


arrie

rCC

chron

iccarrierIQ

Rinterquartile

range





0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)




0

20

40

60

80

100

Ove

rall

surv

ival

()

(c)




0

20

40

60

80

100O

vera

ll su

rviv

al (

)

(d)









0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)





0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)









0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C9

Adjacent Tumor

Plt00001

010002000300040005000600070008000

CYP2

C8

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP2

A6

Adjacent Tumor

Plt00001

minus2000minus1000

010002000300040005000

CYP1

A2

Adjacent Tumor

Plt00001

minus1000minus500

050010001500200025003000

CYP4

A11

Adjacent Tumor

Plt00001

minus2000

0

2000

4000

6000

8000

10000

CYP3

A4

Adjacent Tumor

Plt00001

minus3000

0

3000

6000

9000

12000

15000

CYP2

E1


Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

Adjacent Tumor

Plt00001

0246810121416

CYP1

A2

02468101214161820

CYP2

E1

0246810121416

CYP2

A6

0246810121416

CYP3

A4

02468101214161820

CYP2

C8

02468101214161820

CYP4

A11

0246810121416

CYP2

C9





4 Discussion



Table2Clinicop

atho

logicalcom

paris

ongrou

pedby

CYP2

A6andCY

P2C8

from

GSE

14520andGSE

36376

Category

CYP2

A6

Pvalue

CYP2

C8Pvalue

High

Low

High

Low

GSE

14520

HBV

viralstatus(AV

R-CC

CC

NA)

25814

31745

0635

28784

28775

0943

Alanine

aminotransferase

(gt50lt50U

L)

3872

5159

0099

4268

4763

0583

Maintumor

size(

gt5lt5NAcm)

34760

44651

016

33770

45641

0091

Multin

odular

(yesno)

1892

2783

0181

2189

2486

0738

Cirrho

sis(yesno)

1019

1019

101019

1019

10TN

Msta

ging

(III

IIIN

A)

5737151

3640331

000

55633210

3744272

0032

BCLC

staging

(0-A

BC

NA)

83971

7513

211

0039

79813

07914

152

0357

GSE

36376

Edmon

dson

grade(

IIII

II)1610

04

89715

001

19965

510114

000

2Tu

mor

size(

median)m

m34

(31)

40(45)

0203

34(30)

42(45)

0081

Vascular

invasio

n(yesno)

5169

8436

lt000

15169

8436

lt000

1Major

portalvein

invasio

n(yesno)

1119

8112

003

611

198112

003


(yesno)

1710

33882

000

21810

23783

000

5Multic

entricoccurrence

(yesno)

8112

5115

057

7113

6114

10Dire

ctinvasio

nof

adjacent

organ(yesno)

3117

2118

103117

2118

10AJC

Csta

ging

(1234)

664383

3657252

lt000

1674283

3558252

lt000

1BC

LCsta

ging

(ABC

)75432

64488

0092

80382

59538

000

9Alpha

fetoprotein[M

edian(IQ

R)ngml]

965(822)

2566(41368)

lt000

191

(616

)3038(953445)

lt000

1NAn

otavailableAV

R-CC

activev


arrie

rCC

chron

iccarrierIQ

Rinterquartile

range





0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)




0

20

40

60

80

100

Ove

rall

surv

ival

()

(c)




0

20

40

60

80

100O

vera

ll su

rviv

al (

)

(d)









0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)





0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)









0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of




















Table2Clinicop

atho

logicalcom

paris

ongrou

pedby

CYP2

A6andCY

P2C8

from

GSE

14520andGSE

36376

Category

CYP2

A6

Pvalue

CYP2

C8Pvalue

High

Low

High

Low

GSE

14520

HBV

viralstatus(AV

R-CC

CC

NA)

25814

31745

0635

28784

28775

0943

Alanine

aminotransferase

(gt50lt50U

L)

3872

5159

0099

4268

4763

0583

Maintumor

size(

gt5lt5NAcm)

34760

44651

016

33770

45641

0091

Multin

odular

(yesno)

1892

2783

0181

2189

2486

0738

Cirrho

sis(yesno)

1019

1019

101019

1019

10TN

Msta

ging

(III

IIIN

A)

5737151

3640331

000

55633210

3744272

0032

BCLC

staging

(0-A

BC

NA)

83971

7513

211

0039

79813

07914

152

0357

GSE

36376

Edmon

dson

grade(

IIII

II)1610

04

89715

001

19965

510114

000

2Tu

mor

size(

median)m

m34

(31)

40(45)

0203

34(30)

42(45)

0081

Vascular

invasio

n(yesno)

5169

8436

lt000

15169

8436

lt000

1Major

portalvein

invasio

n(yesno)

1119

8112

003

611

198112

003


(yesno)

1710

33882

000

21810

23783

000

5Multic

entricoccurrence

(yesno)

8112

5115

057

7113

6114

10Dire

ctinvasio

nof

adjacent

organ(yesno)

3117

2118

103117

2118

10AJC

Csta

ging

(1234)

664383

3657252

lt000

1674283

3558252

lt000

1BC

LCsta

ging

(ABC

)75432

64488

0092

80382

59538

000

9Alpha

fetoprotein[M

edian(IQ

R)ngml]

965(822)

2566(41368)

lt000

191

(616

)3038(953445)

lt000

1NAn

otavailableAV

R-CC

activev


arrie

rCC

chron

iccarrierIQ

Rinterquartile

range





0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)




0

20

40

60

80

100

Ove

rall

surv

ival

()

(c)




0

20

40

60

80

100O

vera

ll su

rviv

al (

)

(d)









0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)





0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)









0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of























0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)




0

20

40

60

80

100

Ove

rall

surv

ival

()

(c)




0

20

40

60

80

100O

vera

ll su

rviv

al (

)

(d)









0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)





0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)









0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of























0

20

40

60

80

100

Ove

rall

surv

ival

()

(a)



0

20

40

60

80

100

Ove

rall

surv

ival

()


(b)





0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)




0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(b)









0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of























0

20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)

(a)



20

40

60

80

100

Recu

rren

ce-fr

ee su

rviv

al (

)


(b)





Data Availability


Disclosure




References









































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of





















































of




Disease Markers



OncologyJournal of





PPAR Research



Volume 2018


Journal of

ObesityJournal of



















ReseachArticle - Hindawi Publishing CorporationCYPC, CYPE, CYP A, and CYPA were all dierentially...

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Transcript of ReseachArticle - Hindawi Publishing CorporationCYPC, CYPE, CYP A, and CYPA were all dierentially...