title: [Study of the CpG methylation status of ER alpha gene in estrogen receptor alpha-negative breast cancer cell lines and the role of hydralazine demethylation].
abstract: OBJECTIVE
  To detect the 5'CpG island methylation of estrogen receptor (ER) alpha gene promotor region in ER alpha-negative human breast cancer cell lines (MDA-MB-231 and MDA-MB-435) and breast cancer tissues; and to investigate the possibility of hydralazine in restoring the expression of ER alpha gene through demethylation.
  
  
  METHODS
  The CpG island methylation status of ER alpha gene promotor region-A, B and C in ER alpha-negative human breast cancer cell lines (MDA-MB-231 and MDA-MB-435) and 20 cases of breast cancer tissues were analyzed using methylation-specific polymerase chain reaction (MSP). The mRNA expression profile of ER alpha isoforms (ERalpha-A, ERalpha-B and ERalpha-C) and coding region in ER alpha gene were analyzed by using RT-PCR, after hydralazine treatment.
  
  
  RESULTS
  In the two cell lines studied, CpG island was methylated in ERalpha-A and ERalpha-B but not ERalpha-C. Among them, methylation of CpG island in ERalpha-A was obtained in 13 breast tumor cases; methylation of ERalpha-B seen in 10 tumor cases; methylation of both ERalpha-A and ERalpha-B seen in 9 cases; and methylation of ERalpha-C was obtained in only 1 case. The positivity rates were 65%, 50%, 45% and 5% respectively.
  
  
  CONCLUSIONS
  The ER alpha gene non-expression in breast cancer is probably associated with CpG island methylation in ER alpha gene promotor region A and B, and the level of methylation is enhanced as advance of tumors in clinical stage. Hydralazine, served as a demethylating agent enables to restore the expression of ER alpha gene.
authors.affiliation: Breast Center, Southwest Hospital, Chongqing 400038, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Tang
authors.forename: Jun
authors.initials: J
authors.lastname: Jiang
mesh_terms: D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation; D047628:Estrogen Receptor alpha / Q000096:biosynthesis / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D006830:Hydralazine / Q000494:pharmacology*; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
chemical_list: D047628:Estrogen Receptor alpha; D012333:RNA, Messenger; D006830:Hydralazine