Research Article Prevalence of HER-2 and Hormone Receptors ...

Research ArticlePrevalence of HER-2 and HormoneReceptors and P53 Mutations in the PathologicSpecimens of Breast Cancer Patients

Seyed Abbas Mirmalek1 Maryam Hajilou2 Seyed Alireza Salimi Tabatabaee3

Yekta Parsa3 Soheila Yadollah-Damavandi3 and Tina Parsa3

1 Department of Surgery Islamic Azad University Tehran Medical Sciences Branch Tehran Iran2 International Azad University of Kish Kish Iran3 Studentsrsquo Research Committee Islamic Azad University Tehran Medical Sciences Branch Tehran Iran

Correspondence should be addressed to Yekta Parsa yektaparsagmailcom

Received 20 May 2014 Accepted 24 October 2014 Published 12 November 2014

Academic Editor Vladimir F Semiglazov

Copyright copy 2014 Seyed Abbas Mirmalek et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Prognostic factors are in interest for breast cancer as the second cause of malignancy deaths Some have predictive values as humanepidermal growth factor receptor-2 (HER-2) and estrogen receptor (ER) To access the incidence of HER2 and its relations to otherfactors like age pathology ER progesterone receptor (PR) and P53 2000 pathologic blocks from 2750 total samples have beenselected from 2011 to 2013 in Cancer Institute of Tehran Incidence of HER2 ER PR and P53 was 585 334 433 and 654respectively Invasive ductal carcinoma was the most pathologic type (822) and 60ndash70 positive HER2 and P53 had negativeER and PR (poor prognosis)The peak age of incidence of breast cancer was perimenopausal age group (46ndash55 years) Our cases hadmore positive HER2 and P53 and less positive PR and ER compared to other studies High perimenopausal incidence as anotherfinding assures the importance of breast cancer screening in these age groups

1 Introduction

Among malignancies breast cancer is one of the mostcommon causes of mortality in women Every woman isexposed to 8ndash10 of the risk of this disease during her life [1]

Several prognostic and predictive factors have been usedfor breast malignancies during last yearsThemost importantfactors include P53 progesterone and estrogen receptors andhuman epidermal growth factor receptor-2 (HER-2) HER-2 gene or p185 is a part of the genetic code existing in allhealthy individuals involved in the regulation of normal cellgrowth [2ndash4] If extra copies of this gene are developed onthe cell surface the gene is overexpressed making the celltumoral In general the positive HER-2 receptor has thethree following characteristics rapid tumor growth lowersurvival rate and better response to adjuvant therapies suchas chemotherapy and new drugs such as Herceptin [5ndash7]Therefore understanding the interaction between HER-2

polymorphism and breast cancer risk factors can be effectivein determining treatment strategies and evaluating prognosisin this disease [8]

Increased HER-2 expression is involved in endometrialstomach and prostate cancers as well as breast cancer [9]but they show a lower prevalence of this gene polymorphismcompared to breast cancer [10] There are two methods foridentification of the product of this gene (1) immunohis-tochemistry (IHC) which shows cell surface proteins bystaining with Ab and is more economical and (2) fluo-rescence in situ hybridization (FISH) that is more reliableand conducts the staining at the T-cell level The role ofestrogen in estrogen receptor- (ER-) positive cancer cells isthe development of the cell cycle and prevention of apoptosisTherefore antiestrogens agents stop proliferation during thecell cycle As a mitogen estrogen can generally play therole of a prognostic factor in ER-positive cancer cells Inaddition it is of great value in determining the response

Hindawi Publishing CorporationInternational Journal of Breast CancerVolume 2014 Article ID 564308 3 pageshttpdxdoiorg1011552014564308

2 International Journal of Breast Cancer

to adjuvant therapies such as hormone therapy [7 9ndash11]The presence or absence of progesterone receptor (PR) canalso be of importance for predicting response to hormonaltreatments Despite previous studies simultaneous ERPRpositivity increases the response to endocrine therapies upto 75 On the other hand one-third of ER-positive and PR-negative cases responded to hormone therapies A hypothesissuggests that the goal of the activity of ER is to facilitatethe development of the tumor so it is an effective aspectof this receptor which is being investigated [11] P53 is atranscription factor which is proposed as a tumor suppressorThe mutation of this gene is associated with increased riskof breast cancer and poorer prognosis [12 13] The activityof P53 in tumor suppression via the stoppage of cell cycleor induction results in apoptosis Many experiments haveindicated that besides their higher prevalence p53mutationsare associated with drug resistance [14ndash17] Currently thestaging evaluation that is used for determining the treatmentprocess of patients is different from previous methods Evengrade I PR-negative and ER-negative patients need moreaggressive adjuvant treatments and vice versa Thereforeconsidering the importance of this matter we decided toinvestigate the prevalence of the above factors in breastcancer patients and measure their relationship with eachother and factors such as age and the type of pathology Withregard to the utmost importance of molecular biology inthe diagnosis treatment and even prevention of cancer thepresent research can be a basis for similar studies

2 Materials and Methods

This retrospective cross-sectional study was performed on2000 women whose breast cancer pathology blocks weresent to the Cancer Institute of Imam Khomeini HospitalTehran Iran from 2011 to 2013 All cases whose pathologyresults were one of the types of breast malignancies andalso were investigated for the above factors were selectedfrom about 2750 patients with breast lump whose pathologyblocks were investigated from 2011 to 2013 in this centerThe 2000 present cases in this study were selected from the2750 patients Patients were excluded if they had bilateralbreast cancer untreated brain metastases osteoplastic bonemetastases pleural effusion or ascites as the only evidenceof disease a second type of primary cancer Patients werealso excluded if they were pregnant or had received anytype of therapeutic intervention since their malignancy wasdiagnosed Afterwards the test results of 2000 of them wereextracted from their pathology documents in the archive ofthe Cancer Institute and then recorded All samples wereevaluated by the IHC staining under the direct supervisionof at least two pathology academics HER-2 status was deter-mined by means of IHC using the Dako HercepTest (DakoCopenhagen Denmark) and scored with the Dako scoringsystem [18] Only patients who had weak-to-moderate stain-ing of the entire tumor-cell membrane for Her-2 (referred toas a score of 2+) or more than moderate staining (referred toas a score of 3+) in more than 10 percent of tumor cells onIHC analysis were eligible for the study

Table 1 Frequency of hormone receptors and breast cancer types

Breast cancer types Receptors types Percent

IDC ERPR+ 33ERPRminus 56

DCIS mucinouscarcinoma

HER-2+ 824HER-2minus 846

IDCmedullary carcinoma

P53+ 90P53minus 857

medullary carcinomaILC and IDC

PR+ 857PRminus 56

ER and PR receptors status was determined with a modi-fied avidin-biotin (ABC) immunoperoxidasemethod accord-ing to standard protocols (Vector Laboratories BurlingameCA)The 331015840-diaminobenzidine was used as the chromogenThe immunostaining results for ER and PR were assessedsemiquantitatively and reported as positive if more than 5of cells immunostained in a tumor P53 overexpression wasdefined as more than 50 of the cells with strong nuclearstaining [19]

Collected data are expressed as mean and standarddeviation values All data were analyzed by using ANOVAtest with SPSS software (Version-13) 119875 value lt 005 wasconsidered significant

3 Results and Discussions

The highest rates of breast cancer were observed at the agesof 46ndash55 and lowest rates were observed under the age of25 years and above 66 years The most prevalent malignancywas invasive ductal carcinoma (IDC) at 822 In this group33 and 56 were ERPR-positive and ERPR-negativerespectively Ductal carcinoma in situ (DCIS) and mucinouscarcinoma comprised the highest rates of HER-2-negativeand HER-2-positive cases at 846 and 824 respectivelyInvasive tubular carcinoma (ITC) and medullary carcinomareportedly comprised the highest rates of P53-negative andP53-positive cases at 857 and 90 respectively (Table 1)

The highest rates of PR-negative and PR-positive caseswere observed in medullary and invasive lobular carcinoma(ILC) and ITC at 56 and 857 respectively

The highest rates of positive HER-2 and negative HER-2were observed at the ages of lower than 25 (769) and 26ndash55years (mean age range)The rate of HER-2-positive increasedat the age of over 55 years

The highest and lowest rates of ER-negative and ER-positive were observed at the ages of 56ndash65 and under25 years at 755 and 615 respectively This significantdifference with other studies can be due to sampling bias inthe Cancer Institute The highest rates of PR-negative andPR-positive were observed at the ages of over 66 years andunder 25 years at 611 and 615 respectively There wasno significant relationship between age and P53 (119875 = 0295)(Table 2) It can be noted that 618 of ER-negative cases were

International Journal of Breast Cancer 3

Table 2 Highest and lowest frequency of HER-2 ER and PR statuswithin different age groups

Receptors status Highest age range () Lowest age range ()HER-2+minus lt25 yo (769) 26ndash55 yoER+minus 56ndash65 yo (755) lt25 yo (615)PR+minus gt66 yo (611) lt25 yo (615)

positive regarding HER-2 (poor prognosis) and 481 of ER-positive cases were reported negative regarding HER-2 (goodprognosis)

Additionally 642 of PR-negative cases were positiveregarding HER-2 (poor prognosis) and 49 of PR-positivecases were negative regarding HER-2 (good prognosis)Furthermore 468 of P53-negative cases were negativeregarding HER-2 (good prognosis) and 613 were positiveregarding both factors (poor prognosis)Moreover 844 thecases were negative regarding both factors and 987 of ER-positive cases were PR-positive

In addition 731 of ER-negative cases were P53-positive501 of ER-positive cases were P53-negative 723 of ER-negative cases were P53-positive and 438 of ER-positivecases were P53-negative Interestingly the result obtainedregarding the relationship between HER-2 and the type ofmalignancy and ER and PR had a relationship with thesimilarity of ER and PR receptors in a way that therewas a significant relationship between the above 3 factorsonly if both ER and PR receptors were reported to bepositive or negative Regarding the relationship between ageand the HER-2 receptor poorer prognosis was associatedwith younger ages as expected (with more positive HER-2)However in the others hormone receptor-positive (ER andPR) patients have a higher age range and a good prognosisThis point indicates the need for further research with regardto the lower prevalence of ER and PR in our study

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] R J Pietras J Arboleda D M Reese et al ldquoHER-2 tyro-sine kinase pathway targets estrogen receptor and promoteshormone-independent growth in human breast cancer cellsrdquoOncogene vol 10 no 12 pp 2435ndash2446 1995

[2] K B Horwitz and W L McGuire ldquoSpecific progesteronereceptors in human breast cancerrdquo Steroids vol 25 no 4 pp497ndash505 1975

[3] W L McGuire M De La Garza and G C Chamness ldquoEvalua-tion of estrogen receptor assays in human breast cancer tissuerdquoCancer Research vol 37 no 3 pp 637ndash639 1977

[4] R Hahnel T Woodings and A B Vivian ldquoPrognostic value ofestrogen receptors in primary breast cancerrdquoCancer vol 44 no2 pp 671ndash675 1979

[5] U Bohn J Aguiar C Bilbao et al ldquoPrognostic value of thequantitative measurement of the oncoprotein P185 Her-2neu

in a group of patients with breast cancer and positive nodeinvolvementrdquo International Journal of Cancer vol 101 no 6 pp539ndash544 2002

[6] C DimasM Frangos-Plemenos E Kouskouni and A Kondis-Pafitis ldquoImmunohistochemical study of p185 HER2 and DF

3

in primary breast cancer and correlation with CA-15-3 serumtumor markerrdquo International Journal of Gynecological Cancervol 12 no 1 pp 74ndash79 2002

[7] H K W Koeppen B D Wright A D Burt et al ldquoOverexpres-sion of HER2neu in solid tumours an immunohistochemicalsurveyrdquo Histopathology vol 38 no 2 pp 96ndash104 2001

[8] M Tanner D Gancberg B A D Leo et al ldquoChromogenicin situ hybridization a practical alternative for fluorescence insitu hybridization to detect HER-2neu oncogene amplificationin archival breast cancer samplesrdquo The American Journal ofPathology vol 157 no 5 pp 1467ndash1472 2000

[9] BM Ryan G E Konecny S Kahlert et al ldquoSurvivin expressionin breast cancer predicts clinical outcome and is associatedwithHER2 VEGF urokinase plasminogen activator and PAI-1rdquoAnnals of Oncology vol 17 no 4 pp 597ndash604 2006

[10] M C Ronckers C A Erdmann and C E Land ldquoRadiationand breast cancer a review of current evidencerdquo Breast CancerResearch vol 7 no 1 pp 21ndash32 2005

[11] S Kaptain L K Tan and B Chen ldquoHer-2neu and breastcancerrdquo Diagnostic Molecular Pathology vol 10 no 3 pp 139ndash152 2001

[12] Y Hatanaka K Hashizume Y Kamihara et al ldquoQuantita-tive immunohistochemical evaluation of HER2neu expressionwith HercepTestŮ in breast carcinoma by image analysisrdquoPathology International vol 51 no 1 pp 33ndash36 2001

[13] M A Olayioye ldquoUpdate on HER-2 as a target for cancertherapy intracellular signaling pathways of ErbB2HER-2 andfamily membersrdquo Breast Cancer Research vol 3 no 6 pp 385ndash389 2001

[14] A-M Martin and B L Weber ldquoGenetic and hormonal riskfactors in breast cancerrdquo Journal of the National Cancer Institutevol 92 no 14 pp 1126ndash1135 2000

[15] RM Elledge andD C Allred ldquoThe p53 tumor suppressor genein breast cancerrdquo Breast Cancer Research and Treatment vol 32no 1 pp 39ndash47 1994

[16] D J Slamon W Godolphin L A Jones et al ldquoStudies ofthe HER-2neu proto-oncogene in human breast and ovariancancerrdquo Science vol 244 no 4905 pp 707ndash712 1989

[17] C W Elston and I O Ellis ldquoPathological prognostic factorsin breast cancer I The value of histological grade in breastcancer experience from a large study with long-term follow-uprdquo Histopathology vol 19 no 5 pp 403ndash410 1991

[18] T W Jacobs A M Gown H Yaziji M J Barnes and S JSchnitt ldquoSpecificity of HercepTest in determining HER-2neustatus of breast cancers using the United States Food and DrugAdministration-approved scoring systemrdquo Journal of ClinicalOncology vol 17 no 7 pp 1983ndash1987 1999

[19] D M Barnes E A Dublin C J Fisher D A Levison and RR Millis ldquoImmunohistochemical detection of p53 protein inmammary carcinoma an important new independent indicatorof prognosisrdquo Human Pathology vol 24 no 5 pp 469ndash4761993

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Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

2 International Journal of Breast Cancer

to adjuvant therapies such as hormone therapy [7 9ndash11]The presence or absence of progesterone receptor (PR) canalso be of importance for predicting response to hormonaltreatments Despite previous studies simultaneous ERPRpositivity increases the response to endocrine therapies upto 75 On the other hand one-third of ER-positive and PR-negative cases responded to hormone therapies A hypothesissuggests that the goal of the activity of ER is to facilitatethe development of the tumor so it is an effective aspectof this receptor which is being investigated [11] P53 is atranscription factor which is proposed as a tumor suppressorThe mutation of this gene is associated with increased riskof breast cancer and poorer prognosis [12 13] The activityof P53 in tumor suppression via the stoppage of cell cycleor induction results in apoptosis Many experiments haveindicated that besides their higher prevalence p53mutationsare associated with drug resistance [14ndash17] Currently thestaging evaluation that is used for determining the treatmentprocess of patients is different from previous methods Evengrade I PR-negative and ER-negative patients need moreaggressive adjuvant treatments and vice versa Thereforeconsidering the importance of this matter we decided toinvestigate the prevalence of the above factors in breastcancer patients and measure their relationship with eachother and factors such as age and the type of pathology Withregard to the utmost importance of molecular biology inthe diagnosis treatment and even prevention of cancer thepresent research can be a basis for similar studies

2 Materials and Methods

This retrospective cross-sectional study was performed on2000 women whose breast cancer pathology blocks weresent to the Cancer Institute of Imam Khomeini HospitalTehran Iran from 2011 to 2013 All cases whose pathologyresults were one of the types of breast malignancies andalso were investigated for the above factors were selectedfrom about 2750 patients with breast lump whose pathologyblocks were investigated from 2011 to 2013 in this centerThe 2000 present cases in this study were selected from the2750 patients Patients were excluded if they had bilateralbreast cancer untreated brain metastases osteoplastic bonemetastases pleural effusion or ascites as the only evidenceof disease a second type of primary cancer Patients werealso excluded if they were pregnant or had received anytype of therapeutic intervention since their malignancy wasdiagnosed Afterwards the test results of 2000 of them wereextracted from their pathology documents in the archive ofthe Cancer Institute and then recorded All samples wereevaluated by the IHC staining under the direct supervisionof at least two pathology academics HER-2 status was deter-mined by means of IHC using the Dako HercepTest (DakoCopenhagen Denmark) and scored with the Dako scoringsystem [18] Only patients who had weak-to-moderate stain-ing of the entire tumor-cell membrane for Her-2 (referred toas a score of 2+) or more than moderate staining (referred toas a score of 3+) in more than 10 percent of tumor cells onIHC analysis were eligible for the study

Table 1 Frequency of hormone receptors and breast cancer types

Breast cancer types Receptors types Percent

IDC ERPR+ 33ERPRminus 56

DCIS mucinouscarcinoma

HER-2+ 824HER-2minus 846

IDCmedullary carcinoma

P53+ 90P53minus 857

medullary carcinomaILC and IDC

PR+ 857PRminus 56

ER and PR receptors status was determined with a modi-fied avidin-biotin (ABC) immunoperoxidasemethod accord-ing to standard protocols (Vector Laboratories BurlingameCA)The 331015840-diaminobenzidine was used as the chromogenThe immunostaining results for ER and PR were assessedsemiquantitatively and reported as positive if more than 5of cells immunostained in a tumor P53 overexpression wasdefined as more than 50 of the cells with strong nuclearstaining [19]

Collected data are expressed as mean and standarddeviation values All data were analyzed by using ANOVAtest with SPSS software (Version-13) 119875 value lt 005 wasconsidered significant

3 Results and Discussions

The highest rates of breast cancer were observed at the agesof 46ndash55 and lowest rates were observed under the age of25 years and above 66 years The most prevalent malignancywas invasive ductal carcinoma (IDC) at 822 In this group33 and 56 were ERPR-positive and ERPR-negativerespectively Ductal carcinoma in situ (DCIS) and mucinouscarcinoma comprised the highest rates of HER-2-negativeand HER-2-positive cases at 846 and 824 respectivelyInvasive tubular carcinoma (ITC) and medullary carcinomareportedly comprised the highest rates of P53-negative andP53-positive cases at 857 and 90 respectively (Table 1)

The highest rates of PR-negative and PR-positive caseswere observed in medullary and invasive lobular carcinoma(ILC) and ITC at 56 and 857 respectively

The highest rates of positive HER-2 and negative HER-2were observed at the ages of lower than 25 (769) and 26ndash55years (mean age range)The rate of HER-2-positive increasedat the age of over 55 years

The highest and lowest rates of ER-negative and ER-positive were observed at the ages of 56ndash65 and under25 years at 755 and 615 respectively This significantdifference with other studies can be due to sampling bias inthe Cancer Institute The highest rates of PR-negative andPR-positive were observed at the ages of over 66 years andunder 25 years at 611 and 615 respectively There wasno significant relationship between age and P53 (119875 = 0295)(Table 2) It can be noted that 618 of ER-negative cases were









References








3




















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
























References








3




















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Research Article Prevalence of HER-2 and Hormone Receptors ...

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