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Background  In the present study, we determined the gene hypermethylation profiles of normal tissues adjacent to invasive breast carcinomas and investigated whether these are associated with the gene hypermethylation profiles of the corresponding primary breast tumors.    Methods  A quantitative methylation-specific PCR assay was used to analyze the DNA methylation status of 6 genes ( DAPK, TWIST, HIN-1, RASSF1A, RARβ2  and  APC ) in 9 normal breast tissue samples from unaffected women and in 56 paired cancerous and normal tissue samples from breast cancer patients.    Results  Normal tissue adjacent to breast cancer displayed statistically significant differences to unrelated normal breast tissues regarding the aberrant methylation of the  RASSF1A  (P = 0.03),  RARβ2  (P = 0.04) and  APC  (P = 0.04) genes. Although methylation ratios for all genes in normal tissues from cancer patients were significantly lower than in the cancerous tissue from the same patient (P ≤ 0.01), in general, a clear correlation was observed between methylation ratios measured in both tissue types for all genes tested (P < 0.01). When analyzed as a categorical variable, there was a significant concordance between methylation changes in normal tissues and in the corresponding tumor for all genes tested but  RASSF1A . Notably, in 73% of patients, at least one gene with an identical methylation change in cancerous and normal breast tissues was observed.    Conclusions  Histologically normal breast tissues adjacent to breast tumors frequently exhibit methylation changes in multiple genes. These methylation changes may play a role in the earliest stages of the development of breast neoplasia.

Quantitative methylation profiling in tumor and matched morphologically normal tissues from breast cancer patients