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Estrogen receptor (ER)-positive breast cancers are considered prognostically more favorable than ER-negative tumors, whereas human epidermal growth factor receptor (HER)2/neu-positive breast cancers are associated with worse prognosis. The objective of the present study was to determine whether ER-positive and ER-negative status relates to epigenetic changes in breast cancer-related genes. To evaluate epigenetic differences in tumor-related genes relating to ER and HER2/neu status of primary tumors, we examined the promoter methylation status of the promoter region CpG islands of eight major breast tumor-related genes ( RASSF1A ,  CCND2 ,  GSPT1 ,  TWIST ,  APC ,  NES1 ,  RARβ2 , and  CDH1 ).    Methods  Paired ER-positive ( n  = 65) and ER-negative ( n  = 65) primary breast tumors ( n  = 130) matched for prognostic factors were assessed. DNA was extracted from paraffin-embedded tumor tissue after microdissection, and methylation-specific PCR and capillary-array electrophoresis analysis were performed.    Results  In early stages of tumor progression (T1 and N0),  RASSF1A  and  CCND2  were significantly ( P  < 0.05) more methylated in ER-positive than in ER-negative tumors.  GSTP1  hypermethylation was more frequent in the lymph node metastasis positive group than in the negative group. Double negative (ER-negative, HER2/neu-negative) breast cancers had significantly lesser frequencies of  RASSF1A ,  GSTP1 , and  APC  methylation ( P  < 0.0001,  P  < 0.0001, and  P  = 0.0035, respectively). Both ER and HER2/neu status correlated independently with these epigenetic alterations.    Conclusion  We demonstrated significant differences in tumor-related gene methylation patterns relevant to ER and HER2/neu status of breast tumors. This may be of significance in the assessment of targeted therapy resistance related to ER and HER2/neu status in breast cancer patients.

Estrogen receptor and HER2/neu status affect epigenetic differences of tumor-related genes in primary breast tumors