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Background  Oxidative stress may lead to an increased level of unrepaired cellular DNA damage, which is discussed as one risk for tumor initiation. Mismatch repair (MMR) enzymes act as proofreading complexes that maintain the genomic integrity and MMR-deficient cells show an increased mutation rate. One important gene in the MMR complex is the MutL homolog 1 ( MLH1 ) gene. Since a diet rich in antioxidants has the potential to counteract harmful effects by reactive oxygen species (ROS), we investigated the impact of an antioxidant, folate, and vitamin rich diet on the epigenetic pattern of  MLH1 . These effects were analyzed in individuals with non-insulin depended diabetes mellitus type 2 (NIDDM2) and impaired fasting glucose (IFG).    Methods  In this post-hoc analysis of a randomized trial we analyzed DNA methylation of  MLH1 ,  MSH2 , and  MGMT  at baseline and after 8 weeks of intervention, consisting of 300 g vegetables and 25 ml plant oil rich in polyunsaturated fatty acids per day. DNA methylation was quantified using combined bisulfite restriction enzyme analysis (COBRA) and pyrosequencing.  MLH1  and  DNMT1  mRNA expression were investigated by qRT-PCR. DNA damage was assessed by COMET assay. Student’s two-tailed paired t test and one-way ANOVA with Scheffé corrected Post hoc test was used to determine significant methylation and expression differences. Two-tailed Pearson test was used to determine correlations between methylation level, gene expression, and DNA strand break amount.    Results  The intervention resulted in significantly higher CpG methylation in two particular  MLH1  promoter regions and the  MGMT  promoter. DNA strand breaks and methylation levels correlated significantly. The expression of  MLH1 ,  DNMT1 , and the promoter methylation of  MSH2  remained stable. CpG methylation levels and gene expression did not correlate.    Conclusion  This vitamin and antioxidant rich diet affected the CpG methylation of  MLH1 . The higher methylation might be a result of the ROS scavenging antioxidant rich diet, leading to lower activity of DNA demethylating enzymes. Our results suggest the hypothesis of CpG demethylation via DNA repair enzymes under these circumstances. NIDDM2 and IFG patients benefit from this simple dietary intervention involving epigenetic and DNA repair mechanisms.

Vitamin and antioxidant rich diet increases MLH1 promoter DNA methylation in DMT2 subjects