DNA Methylation at the C-5 Position of Cytosine by Methyl Radicals: A Possible Role for Epigenetic Change during Carcinogenesis by Environmental Agents

During carcinogenesis, methylation of the C-5 position of cytosines in the promoter region of tumor suppressor genes is often observed. Enzymatic DNA methylation is a widely accepted mechanism for this phenomenon. It is interesting to propose a free radical mechanism for 5-methyldeoxycytidine (m(5)dC) production, because the C-5 position of cytosine is an active site for free radical reactions. When deoxycytidine (dC) and cumene hydroperoxide (CuOOH), a tumor promoter and a methyl radical producer, were reacted in the presence of ferrous ion at pH 7.4, the formation of m(5)dC was observed. The same reaction also proceeded with t-butyl hydroperoxide (BuOOH). The formation of m(5)dC was also observed in DNA by the CuOOH treatment. This is the first report of chemical DNA methylation at cytosine C-5 by environmental tumor promoters. We propose here that this reaction is one of the important mechanisms of de novo DNA methylation during carcinogenesis, because methyl radicals are produced by the biotransformation of various endogenous and exogenous compounds.