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Background
It has been estimated that approximately 30% of all cancer cases are attributable to diet and lifestyle. One of the mechanisms by which diet may affect cancer risk is through epigenetic processes such as DNA methylation. Folate, and its synthetic form folic acid, function as donor of one-carbon units and have been implicated in regulation of DNA methylation as well as DNA synthesis.
Epidemiological studies suggest that suboptimal levels of folate and other B-vitamins may affect DNA methylation and thereby influence genomic stability and cancer risk, although the exact underlying mechanisms have not been clarified.
Aim
The aim of this project was to identify the effects of long-term supplementation with physiological doses of folic acid and vitamin B12 on global and genome-wide DNA methylation in elderly subjects.
Methods
The current project was implemented as part of a randomised and placebo-controlled trial on the effects of supplemental intake of folic acid and vitamin B12 on fracture incidence. A total of 2919 subjects with mildly elevated homocysteine concentrations (≥12 μM), aged 65 years and older, participated in this study.
Participants were randomly assigned to take 400 μg folic acid and 500 μg vitamin B12 per day or a placebo during an intervention period of two years. Blood was collected at baseline and after two years of intervention. DNA was isolated from buffy coats and bisulfite converted.
For the current project, global and genome-wide DNA methylation were determined for 74 and 89 participants, respectively. The LINE-1 assay was used to assess global DNA methylation, whereas DNA methylation arrays were used to study genome-wide DNA methylation.
These arrays allow determination of the DNA methylation status of more than 450,000 sites, also called CpG sites, simultaneously.
Results
The findings of our study show that long-term supplementation with folic acid and vitamin B12 did not result in pronounced changes in global or genome-wide DNA methylation as compared to the placebo group. Changes in global DNA methylation in participants receiving folic acid and vitamin B12 did not statistically significantly differ from DNA methylation changes in participants receiving a placebo.
Genome-wide analysis of DNA methylation revealed that after the intervention with folic acid and vitamin B12, 33 of the 451,047 CpG sites were significantly differentially methylated as compared to baseline (p<1E-05). Likewise, in the placebo group, 13 CpG sites were significantly differentially methylated as compared to baseline.
Conclusions
In conclusion, this project showed that long-term supplementation with folic acid and vitamin B12 did not have pronounced effects on global or genome-wide DNA methylation in elderly subjects with mildly elevated homocysteine levels.