Tumor Biology

, Volume 33, Issue 2, pp 413–420 | Cite as

Promoter methylation status and expression of estrogen receptor alpha in familial breast cancer patients

  • Jing Wei
  • Bing Han
  • Xiao-yun Mao
  • Min-jie Wei
  • Fan Yao
  • Feng Jin
Research Article


The hypermethylation of estrogen receptor alpha (ERα) promoter is a common molecular alteration in sporadic breast cancer (BC), but its involvement in familial BC remains largely unknown. In the present study, we analyzed the methylation statuses of four regions (ER1, ER3, ER4, and ER5) of the ERα promoter and the ERα expression levels of 113 familial BC patients in a Han Chinese Population from northeastern China and evaluated the association between major clinicopathological features and the hypermethylation statuses of the ERα gene. Tumor samples were analyzed for ERα methylation status by the methylation-specific polymerase chain reaction for ERα, PR, p53, BRCA-1, and BRCA-2 by immunohistochemical (IHC) staining and for Her-2 status by IHC and fluorescence in situ hybridization (FISH). ERα methylation was observed in tumor tissues in 47/113 (41.6%) familial BC patients. There were no significant differences in the methylation statuses among ER1 (20.4%), ER3 (18.6%), ER4 (17.7%), and ER5 (19.5%; χ 2 = 3.89, p > 0.05). An association between ERα expression level and its promoter methylation level was found. In addition, ERα methylation was significantly correlated with tumor size, PR expression, p53 nuclear accumulation, and BRCA-1 and BRCA-2 statuses. In conclusion, in familial BC patients, the level of ERα gene promoter methylation correlates with ERα expression, PR, p53 nuclear accumulation, and BRCA-1 and BRCA-2 statuses. Epigenetic alteration of ERα gene may play an important role in the pathogenesis of familial BC.


ERα Methylation Familial breast cancer Chinese women 


Conflicts of interest



  1. 1.
    Ahmad Z, Khurshid A, Qureshi A, Idress R, Asghar N, Kayani N. Breast carcinoma grading, estimation of tumor size, axillary lymph node status, staging, and nottingham prognostic index scoring on mastectomy specimens. Indian J Pathol Microbiol. 2009;52:477–81.PubMedCrossRefGoogle Scholar
  2. 2.
    Narod SA. Genes, the environment, and breast cancer. Lancet. 2010;375:2123–4.PubMedCrossRefGoogle Scholar
  3. 3.
    Pharoah PD, Day NE, Duffy S, Easton DF, Ponder BA. Family history and the risk of breast cancer: a systematic review and meta-analysis. Int J Cancer. 1997;71:800–9.PubMedCrossRefGoogle Scholar
  4. 4.
    Duss S, André S, Nicoulaz AL, Fiche M, Bonnefoi H, Brisken C, et al. An oestrogen-dependent model of breast cancer created by transformation of normal human mammary epithelial cells. Breast Cancer Res. 2007;9:R38.PubMedCrossRefGoogle Scholar
  5. 5.
    Dunnwald LK, Rossing MA, Li CI. Hormone receptor status, tumor characteristics, and prognosis: a prospective cohort of breast cancer patients. Breast Cancer Res. 2007;9:R6.PubMedCrossRefGoogle Scholar
  6. 6.
    Jones PA, Baylin SB. The fundamental role of epigenetic events in cancer. Nat Rev Genet. 2002;3:415–28.PubMedCrossRefGoogle Scholar
  7. 7.
    Brinkman JA, El-Ashry D. ER. re-expression and re-sensitization to endocrine therapies in ER-negative breast cancers. J Mammary Gland Biol Neoplasia. 2009;14:67–78.PubMedCrossRefGoogle Scholar
  8. 8.
    Flanagan J, Kugler S, Waddell N, Johnstone C, Marsh A, Henderson S, et al. DNA methylome of familial breast cancer identifies distinct profiles defined by mutation status. Breast Cancer Res. 2010;12 Suppl 1:O4.PubMedCrossRefGoogle Scholar
  9. 9.
    Yoshida T, Eguchi H, Nakachi K, Tanimoto K, Higashi Y, Suemasu K, et al. Distinct mechanisms of loss of estrogen receptor alpha gene expression in human breast cancer: methylation of the gene and alteration of trans-acting factors. Carcinogenesis. 2000;21:2193–201.PubMedCrossRefGoogle Scholar
  10. 10.
    Wirtenberger M, Tchatchou S, Hemminki K, Schmutzhard J, Sutter C, Schmutzler RK, et al. Associations of genetic variants in the estrogen receptor coactivators PPARGC1A, PPARGC1B and EP300 with familial breast cancer. Carcinogenesis. 2006;27:2201–8.PubMedCrossRefGoogle Scholar
  11. 11.
    Wirtenberger M, Schmutzhard J, Hemminki K, Meindl A, Sutter C, Schmutzler RK, et al. The functional genetic variant Ile646Val located in the kinase binding domain of the A-kinase anchoring protein 10 is associated with familial breast cancer. Carcinogenesis. 2007;28:423–6.PubMedCrossRefGoogle Scholar
  12. 12.
    Tavassoli FA. Breast pathology: rationale for adopting the ductal intraepithelial neoplasia (DIN) classification. Nat Clin Pract Oncol. 2005;2:116–7.PubMedCrossRefGoogle Scholar
  13. 13.
    Zhao L, Wang L, Jin F, Ma W, Ren J, Wen X, et al. Silencing of estrogen receptor alpha (ERalpha) gene by promoter hypermethylation is a frequent event in Chinese women with sporadic breast cancer. Breast Cancer Res Treat. 2009;117:253–9.PubMedCrossRefGoogle Scholar
  14. 14.
    Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA. 1996;93:9821–6.PubMedCrossRefGoogle Scholar
  15. 15.
    Lapidus RG, Nass SJ, Butash KA, Parl FF, Weitzman SA, Graff JG, et al. Mapping of ER gene CpG island methylation-specific polymerase chain reaction. Cancer Res. 1998;58:2515–9.PubMedGoogle Scholar
  16. 16.
    Kok LF, Lee MY, Tyan YS, Wu TS, Cheng YW, Kung MF, et al. Comparing the scoring mechanisms of p16INK4a immunohistochemistry based on independent nucleic stains and independent cytoplasmic stains in distinguishing between endocervical and endometrial adenocarcinomas in a tissue microarray study. Arch Gynecol Obstet. 2010;281:293–300.PubMedCrossRefGoogle Scholar
  17. 17.
    Koo CL, Kok LF, Lee MY, Wu TS, Cheng YW, Hsu JD, et al. Scoring mechanisms of p16INK4a immunohistochemistry based on either independent nucleic stain or mixed cytoplasmic with nucleic expression can significantly signal to distinguish between endocervical and endometrial adenocarcinomas in a tissue microarray study. J Transl Med. 2009;7:25.PubMedCrossRefGoogle Scholar
  18. 18.
    Hatanaka Y, Hashizume K, Kamihara Y, Itoh H, Tsuda H, Osamura RY, et al. Quantitative IHC evaluation of HER2/neu expression with HercepTest™ in breast carcinoma by image analysis. Pathol Int. 2001;51:33–6.PubMedCrossRefGoogle Scholar
  19. 19.
    Hoff ER, Tubbs RR, Myles JL, Procop GW. HER2/neu amplification in breast cancer: stratification by tumor type and grade. Am J Clin Pathol. 2002;117:916–21.PubMedCrossRefGoogle Scholar
  20. 20.
    Murff HJ, Byrne D, Haas JS, Puopolo AL, Brennan TA. Race and family history assessment for breast cancer. J Gen Intern Med. 2005;20:75–80.PubMedCrossRefGoogle Scholar
  21. 21.
    Yehiely F, Moyano JV, Evans JR, Nielsen TO, Cryns VL. Deconstructing the molecular portrait of basal-like breast cancer. Trends Mol Med. 2006;12:537–44.PubMedCrossRefGoogle Scholar
  22. 22.
    Pirouzpanah S, Taleban FA, Atri M, Abadi AR, Mehdipour P. The effect of modifiable potentials on hypermethylation status of retinoic acid receptor-beta2 and estrogen receptor-alpha genes in primary breast cancer. Cancer Causes Control. 2010;21:2101–11.PubMedCrossRefGoogle Scholar
  23. 23.
    Mirza S, Sharma G, Prasad CP, Parshad R, Srivastava A, Gupta SD, et al. Promoter hypermethylation of TMS1, BRCA1, ERalpha and PRB in serum and tumor DNA of invasive ductal breast carcinoma patients. Life Sci. 2007;81:280–7.PubMedCrossRefGoogle Scholar
  24. 24.
    Nass SJ, Herman JG, Gabrielson E, Iversen PW, Parl FF, Davidson NE, et al. Aberrant methylation of the estrogen receptor and E-cadherin 5′ CpG islands increases with malignant progression in human breast cancer. Cancer Res. 2000;60:4346–8.PubMedGoogle Scholar
  25. 25.
    Hockings JK, Thorne PA, Kemp MQ, Morgan SS, Selmin O, Romagnolo DF. The ligand status of the aromatic hydrocarbon receptor modulates transcriptional activation of BRCA-1 promoter by estrogen. Cancer Res. 2006;66:2224–32.PubMedCrossRefGoogle Scholar
  26. 26.
    Burkadze G, Khardzeishvili O, Gudadze M, Tsikhiseli G, Turashvili G. Immunohistochemical expression of BRCA1 protein in invasive ductal carcinoma of the breast. Georgian Med News. 2010;184–185:51–60.PubMedGoogle Scholar
  27. 27.
    Flanagan J, Kugler S, Waddell N, Johnstone C, Marsh A, Henderson S, et al. DNA methylome of familial breast cancer identifies distinct profiles defined by mutation status. Am J Hum Genet. 2010;86:420–33.PubMedCrossRefGoogle Scholar
  28. 28.
    Hosey AM, Gorski JJ, Murray MM, Quinn JE, Chung WY, Stewart GE, et al. Molecular basis for estrogen receptor alpha deficiency in BRCA1-linked breast cancer. J Natl Cancer Inst. 2007;99:1683–94.PubMedCrossRefGoogle Scholar
  29. 29.
    Huang YS, Zhi YF, Wang SR. Hypermethylation of estrogen receptor-alpha gene in atheromatosis patients and its correlation with homocysteine. Pathophysiology. 2009;16:259–65.PubMedCrossRefGoogle Scholar
  30. 30.
    Asada H, Yamagata Y, Taketani T, Matsuoka A, Tamura H, Hattori N, et al. Potential link between estrogen receptor-alpha gene hypomethylation and uterine fibroid formation. Mol Hum Reprod. 2008;14:539–45.PubMedCrossRefGoogle Scholar
  31. 31.
    Roos MA, Bock GH, Vries J, Vegt B, Wesseling J. p53 overexpression is a predictor of local recurrence after treatment for both in situ and invasive ductal carcinoma of the breast. J Surg Res. 2007;140:109–14.PubMedCrossRefGoogle Scholar
  32. 32.
    Rossner Jr P, Gammon MD, Zhang YJ, Terry MB, Hibshoosh H, Memeo L, et al. Mutations in p53, p53 protein overexpression and breast cancer survival. J Cell Mol Med. 2009;13:3847–57.PubMedCrossRefGoogle Scholar
  33. 33.
    Sayeed A, Konduri SD, Liu W, Bansal S, Li F, Das GM. Estrogen receptor alpha inhibits p53-mediated transcriptional repression: implications for the regulation of apoptosis. Cancer Res. 2007;67:7746–55.PubMedCrossRefGoogle Scholar
  34. 34.
    Shirley SH, Rundhaug JE, Tian J, Cullinan-Ammann N, Lambertz I, Conti CJ, et al. Transcriptional regulation of estrogen receptor-alpha by p53 in human breast cancer cells. Cancer Res. 2009;69:3405–14.PubMedCrossRefGoogle Scholar
  35. 35.
    Mao XY, Fan CF, Zheng HC, Wei J, Yao F, Jin F. p53 nuclear accumulation and ERalpha expression in ductal hyperplasia of breast in a cohort of 215 Chinese women. J Exp Clin Cancer Res. 2010;29:112.PubMedCrossRefGoogle Scholar
  36. 36.
    Alsner J, Jensen V, Kyndi M, Offersen BV, Vu P, Børresen-Dale AL, et al. A comparison between p53 accumulation determined by immunohistochemistry and TP53 mutations as prognostic variables in tumours from breast cancer patients. Acta Oncol. 2008;47:600–7.PubMedCrossRefGoogle Scholar
  37. 37.
    Gasco M, Yulug IG, Crook T. TP53 mutations in familial breast cancer: functional aspects. Hum Mutat. 2003;21:301–6.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2011

Authors and Affiliations

  • Jing Wei
    • 1
  • Bing Han
    • 2
  • Xiao-yun Mao
    • 1
  • Min-jie Wei
    • 3
  • Fan Yao
    • 1
  • Feng Jin
    • 1
  1. 1.Departments of Breast Surgery and Surgical Oncology, Research Unit of General SurgeryThe First Affiliated Hospital of China Medical UniversityShenyangChina
  2. 2.Department of First Minimal Invasive Surgery and Bile Duct and Vascular SurgerySheng Jing Hospital of China Medical UniversityShenyangChina
  3. 3.Department of PharmacologyChina Medical UniversityShenyangChina

Personalised recommendations