title: Promoter hypermethylation of MGMT is associated with protein loss in gastric carcinoma.
abstract: Aberrant methylation of CpG islands within promoter regions is associated with transcriptional inactivation of various tumor suppressor genes in neoplasms. Recently, O(6)-methylguanine-DNA methyltransferase, MGMT, was shown to be hypermethylated in certain carcinomas, resulting in loss of MGMT protein. We studied DNA methylation of CpG islands of the MGMT gene by methylation specific PCR in 26 gastric carcinoma tissues and 8 gastric carcinoma cell lines for comparison with levels of MGMT protein expression. In addition, we examined p53 mutation status in the same tissues by PCR-SSCP analysis for comparison with MGMT protein expression levels. In total, promoter hypermethylation of the MGMT gene was found in 8 (31%) of the 26 gastric carcinomas with reduced expression of MGMT protein, whereas the hypermethylation was not detected in the 18 carcinomas with non-reduced MGMT expression. MGMT protein expression levels were associated with promoter hypermethylation of MGMT (p = 0.0001; Mann-Whitney test); however, MGMT expression was not associated with p53 mutation status (p = 0.461; Mann-Whitney test). Among in gastric carcinoma cell lines, the TMK-1 cell line showed loss of the MGMT protein association with promoter hypermethylation and this loss was rectified by treatment with a demethylating agent, 5-Aza-2'-deoxycytidine. Our results suggest that transcriptional inactivation of MGMT by aberrant methylation of the promoter region may participate in carcinogenesis in the stomach.
authors.affiliation: First Department of Pathology, Hiroshima University School of Medicine, Hiroshima, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Oue
authors.forename: H
authors.initials: H
authors.lastname: Shigeishi
authors.forename: H
authors.initials: H
authors.lastname: Kuniyasu
authors.forename: H
authors.initials: H
authors.lastname: Yokozaki
authors.forename: K
authors.initials: K
authors.lastname: Kuraoka
authors.forename: R
authors.initials: R
authors.lastname: Ito
authors.forename: W
authors.initials: W
authors.lastname: Yasui
mesh_terms: D000483:Alleles; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D015153:Blotting, Western; D002277:Carcinoma / Q000235:genetics*; D018899:CpG Islands; D019175:DNA Methylation; D000077209:Decitabine; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016158:Genes, p53 / Q000235:genetics; D006801:Humans; D009154:Mutation; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000235:genetics*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D000964:Antimetabolites, Antineoplastic; D004791:Enzyme Inhibitors; D000077209:Decitabine; D019853:O(6)-Methylguanine-DNA Methyltransferase; D001374:Azacitidine