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DNA methylation and one-carbon metabolism related nutrients and polymorphisms: analysis after mandatory flour fortification with folic acid

  • Josiane Steluti (a1), Cecília Zanin Palchetti (a1), Andreia Machado Miranda (a1), Regina Mara Fisberg (a1) and Dirce Maria Marchioni (a1)...


There is a growing research interest in determining whether changes in the global status of DNA methylation are related to the environment, in particular, to one-carbon metabolism. So, our aim was to investigate the effect of dietary methyl-group donor intake (methionine, folate, choline, betaine, vitamins B2, B6 and B12), biomarkers (total folate, unmetabolised folic acid (FA), 5-methyltetrahydrofolate, homocysteine, vitamins B6 and B12 concentrations) and genetic variants (polymorphisms involved in one-carbon metabolism) on global DNA methylation in a population exposed to mandatory flour fortification with FA. A cross-sectional study of health and living conditions was conducted among a representative sample of residents in São Paulo, Brazil. The mean of global DNA methylation was lower in young people than in adults and the elderly (P = 0·049). No differences between genotypes of polymorphism and global DNA methylation mean were identified. We observed that the increase in betaine intake led to an absolute change in percentage of DNA methylation (β = 0·0005, P = 0·024) using multiple regression. Betaine intake alone was associated with an absolute change in percentage of global DNA methylation. The study did not find an association between global DNA methylation and folate status even in a population exposed to mandatory flour fortification with FA.


Corresponding author

*Corresponding author: Josiane Steluti, fax +55 11 3061 7705, email


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1.Jin, B, Li, Y & Robertson, KD (2011) DNA methylation: superior or subordinate in the epigenetic hierarchy? Genes Cancer 2, 607617.
2.Pauwels, S, Ghosh, M, Duca, RC, et al. (2016) Dietary and supplemental maternal methyl-group donor intake and cord blood DNA methylation. Epigenetics 12, 110.
3.Deng, G, Nguyen, A, Tanaka, H, et al. (2006) Regional hypermethylation and global hypomethylation are associated with altered chromatin conformation and histone acetylation in colorectal cancer. Int J Cancer 118, 29993005.
4.Merlo, A, Herman, JG, Mao, L, et al. (1995) 5' CpG island methylation is associated with transcriptional silencing of the tumour suppressorp16/CDKN2/MTS1 in human cancers. Nat Med 1, 686692.
5.Nelson, HH, Marsit, CJ & Kelsey, KT (2011) Global methylation in exposure biology and translational medical science. Environ Health Perspect 119, 15281533.
6.Brunaud, L, Alberto, JM, Ayav, A, et al. (2003) Effects of vitamin B12 and folate deficiencies on DNA methylation and carcinogenesis in rat liver. Clin Chem Lab Med 41, 10121019.
7.Anderson, OS, Sant, KE & Dolinoy, DC (2012) Nutrition and epigenetics: an interplay of dietary methyl donors, one-carbon metabolism and DNA methylation. J Nutr Biochem 23, 853859.
8.DeVos, L, Chanson, A, Liu, Z, et al. (2008) Associations between single nucleotide polymorphisms in folate uptake and metabolizing genes with blood folate, homocysteine, and DNA uracil concentrations. Am J Clin Nutr 88, 11491158.
9.Crider, KS, Yang, TP, Berry, RJ, et al. (2012) Folate and DNA methylation: a review of molecular mechanisms and the evidence for folate’s role. Adv Nutr 3, 2138.
10.Food Fortification Initiative (2016) Global experiences and evidence for fortifying with folic acid. (accessed November 2018).
11.Giovannucci, E (2002) Epidemiologic studies of folate and colorectal neoplasia: a review. J Nutr 132, 2350S2355S.
12.Cole, BF, Baron, JA, Sandler, RS, et al. (2007) Folic acid for the prevention of colorectal adenomas: a randomized clinical trial. JAMA 6, 23512359.
13.Kim, DH, Smith-Warner, SA, Spiegelman, D, et al. (2010) Pooled analyses of 13 prospective cohort studies on folate intake and colon cancer. Cancer Causes Control 21, 19191930.
14.Kim, MS, Lee, J & Sidransky, D (2010) DNA methylation markers in colorectal cancer. Cancer Metastasis Rev 29, 181206. Carvalho, AM, César, CL, Fisberg, RM, et al. (2014) Meat consumption in São Paulo-Brazil: trend in the last decade. PLOS ONE 9, e96667.
16.Instituto Brasileiro de Geografia e Estatística – IBGE (2005) Pesquisa Nacional por Amostra de Domicílios (National Household Sample Survey). (accessed November 2018).
17.Instituto Brasileiro de Geografia e Estatística – IBGE (2013) Características Étnico-raciais da População Classificações e identidades. Indicadores Sociais Estudos e Análises Informação Demográfica e Socioeconômica número 2 (Ethnic-Racial Characteristics of Population and Identities. Social Studies Indicators and Analysis of Demographic and Socioeconomic Information Number 2). Rio de Janeiro. (accessed July 2019).
18.Guenther, PM, DeMaio, TJ, Ingwersen, LA, et al. (1995) The multiple-pass approach for the 24 hour recall in the Continuing Survey of Food Intakes by Individuals (CSFII) 1994–1996. Paper presented at the International Conference on Dietary Assessment Methods. Boston, MA.
19.Blanton, CA, Moshfegh, AJ, Baer, DJ, et al. (2006) The USDA automated multiple-pass method accurately estimates group total energy and nutrient intake. J Nutr 136, 25942599.
20.Universidade de São Paulo/Faculdade de Ciências Farmacêuticas/Departamento de Alimentos e Nutrição Experimental/BRASILFOODS (2008) Tabela Brasileira de Composição de Alimentos-USP. Versão 5.0 (Brazilian Table of Food Composition-USP. Version 5.0). (accessed November 2018).
21.Kalmbach, R, Paul, L & Selhub, J (2011) Determination of unmetabolized folic acid in human plasma using affinity HPLC. Am J Clin Nutr 94, S343S347.
22.Rybak, ME, Jain, RB & Pfeiffer, CM (2005) Clinical vitamin B6 analysis: an interlaboratory comparison of pyridoxal 5'-phosphate measurements in serum. Clin Chem 51, 12231231.
23.Beckman Coulter Inc. (2009) Vitamin B12 – Access Immunoassay Systems®. California, Technical Lab Manual.
24.Abbott Diagnostics Division (2011) Homocysteine – Abbott Architect i2000. United Kingdom, Technical Lab Manual.
25.Miller, SA, Dykes, DD & Polesky, HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16, 1215.
26.Myakishev, MV, Khripin, Y, Hu, S, et al. (2001) High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers. Genome Res 11, 163169. Oliveira Andrade, F, Fontelles, CC, Rosim, MP, et al. (2014) Exposure to lard-based high-fat diet during fetal and lactation periods modifies breast cancer susceptibility in adulthood in rats. J Nutr Biochem 25, 613622.
28.Davis, CD & Uthus, EO (2004) DNA methylation, cancer susceptibility, and nutrient interactions. Exp Biol Med (Maywood) 229, 988995.
29.Anderson, OS, Sant, KE & Dolinoy, DC (2012) Nutrition and epigenetics: an interplay of dietary methyl donors, one-carbon metabolism and DNA methylation. J Nutr Biochem 23, 853859.
30.Field, MS & Stover, PJ (2017) Safety of folic acid. Ann N Y Acad Sci 1414, 5971.
31.Bailey, SW & Ayling, JE (2009) The extremely slow and variable activity of dihydrofolate reductase in human liver and its implications for high folic acid intake. Proc Natl Acad Sci U S A 106, 1542415429.
32.Steluti, J, Reginaldo, C, Selhub, J, et al. (2019) Presence of circulating folic acid in plasma and its relation with dietary intake, vitamin B complex concentrations and genetic variants. Eur J Nutr 58, 30693077.
33.Linhart, HG, Troen, A, Bell, GW, et al. (2008) Folate deficiency induces genomic uracil misincorporation and hypomethylation but does not increase DNA point mutations. Gastroenterology 136, 227235.e3.
34.Charles, MA, Johnson, IT & Belshaw, NJ (2012) Supra-physiological folic acid concentrations induce aberrant DNA methylation in normal human cells in vitro. Epigenetics 7, 689694.
35.Bae, S, Ulrich, CM, Bailey, LB, et al. (2013) Impact of folic acid fortification on global DNA methylation and one-carbon biomarkers in the Women’s Health Initiative Observational Study cohort. Epigenetics 9, 396403.
36.Obeid, R & Herrmann, W (2012) The emerging role of unmetabolized folic acid in human diseases: myth or reality? Curr Drug Metab 13, 11841195.
37.Pfeiffer, CM, Sternberg, MR, Fazili, Z, et al. (2015) Unmetabolized folic acid is detected in nearly all serum samples from US children, adolescents, and adults. J Nutr 145, 520531.
38.Chamberlain, JA, Dugué, PA, Bassett, JK, et al. (2018) Dietary intake of one-carbon metabolism nutrients and DNA methylation in peripheral blood. Am J Clin Nutr 108, 611621.
39.Kok, DE, Dhonukshe-Rutten, RA, Lute, C, et al. (2015) The effects of long-term daily folic acid and vitamin B12 supplementation on genome-wide DNA methylation in elderly subjects. Clin Epigenetics 7, 121.
40.Dhana, A, Yen, H, Li, T, et al. (2018) Intake of folate and other nutrients related to one-carbon metabolism and risk of cutaneous melanoma among US women and men. Cancer Epidemiol 55, 176183.
41.Stern, LL, Mason, JB, Selhub, J, et al. (2000) Genomic DNA hypomethylation, a characteristic of most cancers, is present in peripheral leukocytes of individuals who are homozygous for the C677T polymorphism in the methylenetetrahydrofolate reductase gene. Cancer Epidemiol Biomarkers Prev 9, 849853.
42.Friso, S, Girelli, D, Trabetti, E, et al. (2005) The MTHFR 1298A>C polymorphism and genomic DNA methylation in human lymphocytes. Cancer Epidemiol Biomarkers Prev 14, 938943.
43.Farkas, SA, Befekadu, R, Hahn-Strömberg, V, et al. (2015) DNA methylation and expression of the folate transporter genes in colorectal cancer. Tumour Biol 36, 55815590.
44.Choi, SW & Mason, JB (2000) Folate and carcinogenesis: an integrated scheme. J Nutr 130, 129132.
45.Kurdyukov, S & Bullock, M (2016) DNA methylation analysis: choosing the right method. Biology (Basel) 5, E3.
46.Wu, HC, Delgado-Cruzata, L, Flom, JD, et al. (2011) Global methylation profiles in DNA from different blood cell types. Epigenetics 6, 7685.


DNA methylation and one-carbon metabolism related nutrients and polymorphisms: analysis after mandatory flour fortification with folic acid

  • Josiane Steluti (a1), Cecília Zanin Palchetti (a1), Andreia Machado Miranda (a1), Regina Mara Fisberg (a1) and Dirce Maria Marchioni (a1)...


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