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Nuovo, Gerard J; Nakagawa, Hidewaki; Sotamaa, Kaisa; Chapelle, Albert de la
Diagnostic molecular pathology 15, Issue: 1Journal Article
Hypermethylation of the MLH1 promoter is present in most sporadic colorectal cancers and leads to the abrogation of MLH1 transcription. The significance of the hypermethylation of the promoter that can be detected in rare foci of cells in the normal colonic epithelium adjacent to the carcinoma is unclear. The purpose of this study was to determine the distribution of hypermethylation of the MLH1 promoter DNA with MLH1 RNA expression by RT in situ PCR and MLH1 protein expression by immunohistochemistry in normal colonic tissues. Hypermethylation of the MLH1 promoter DNA was not evident in sections of normal colon from 10 people with no history of colon cancer. MLH1 RNA and protein were detected diffusely throughout the colonic epithelium in these 10 controls. In comparison, hypermethylation of either of 2 regions of the promoter was evident in the normal mucosa from cases adjacent to adenocarcinoma of the colon. In situ analysis of the DNA methylation showed that there were rare foci that involved small groups of cells, typically toward the middle and surface area of a given crypt. In these areas, small foci of colonic epithelial cells, typically in groups of 5 to 10, were found to lack MLH1 RNA and protein in adjacent serial sections, consistent with silencing of the gene by hypermethylation that may play a role in the origin of these neoplasms. It is concluded that the rare foci of hypermethylation of the MLH1 promoter in normal colon epithelium from people with colorectal cancer is associated with silencing of the gene that may play a role in the early evolution of these neoplasms. A combination of methylation-specific in situ PCR with in situ RNA and protein analysis can assist in documenting rare foci of promoter hypermethylation-induced gene silencing.
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