Abstract
The development of certain human cancers has been linked with inadequate intake of folates. The effects of folate deficiency in vivo on DNA stability (strand breakage, misincorporated uracil and oxidative base damage) in lymphocytes isolated from rats fed a diet deficient in folic acid was determined. Because the metabolic pathways of folate and other methyl donors are closely coupled, the effects of methionine and choline deficiency alone or in combination with folate deficiency were determined. Feeding male Hooded Lister rats a folate-free diet for 10 weeks created a moderate folate deficiency (25–50% (approx.) decrease in plasma, red blood cell and hepatic folate concentrations (P< 0.05) and a 20% rise in plasma homocysteine (P<0.05). Lymphocyte DNA strand breakage was increased successively in all groups after 4 weeks and 8 weeks on the diet (50–100% (approx.) after 8 weeks). Only low folate specifically and progressively induced uracil misincorporation throughout the study (100% (approx.) after 8 weeks). Neither folate deficiency nor choline/methionine deficiency altered oxidative DNA base damage. In summary, moderate folate deficiency in vivo is associated with a decrease in DNA stability, measured as increased DNA strand breakage and misincorporated uracil. © 2000 Cancer Research Campaign http://www.bjcancer.com
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Albright CD, Liu R, Mar MH, Shin OH, Vrablic AS, Salganik RI and Zeisel SH (1997 Diet, apoptosis and carcinogenesis. Adv Exp Med Biol 422: 97–107
Asahara H, Wistort PM, Bank JF, Bakerian RH and Cunningham RP (1989 Purification and characterisation of Escherichia coli endonuclease III from the cloned n th gene. Biochemistry 28: 4444–4449
Bailey LB (1995 Folate requirements and dietary recommendations. Bailey LB (eds), Folate in Health and Disease, Marcel Dekker, Inc: New York pp 123–151
Blount BC and Ames BN (1994 Analysis of uracil in DNA by gas chromatography–mass spectrometry. Anal Biochem 219: 195–200
Blount BC, Mack MM, Wehr CM, MacGregor JT, Hiatt RA, Wang G, Wickramasinghe SN, Everson RB and Ames BN (1997 Folate deficiency causes uracil misincorporation into human DNA and chromosomal breakage: implications for cancer and neuronal damage. Proc Natl Acad Sci USA 94: 3290–3295
Czeizel AE (1993 Prevention of congenital abnormalities by periconceptional multivitamin supplementation. BMJ 306: 1645–1649
DeBree A, van Dusseldorp M, Brouwer IA, van het Hof KH and Steegers-Theunissen RPM (1997 Review: Folate intake in Europe: recommended, actual and desired intake. Eur J Clin Nutr 51: 643–660
Duthie SJ (1999 Folic acid deficiency and cancer: mechanisms of DNA instability. Brit Med Bull 55: 578–592
Duthie SJ and Hawdon A (1998 DNA stability (strand breakage, uracil misincorporation, and defective repair) is increased by folic acid depletion in human lymphocytes in vitro. FASEB J 12: 1491–1497
Duthie SJ, Ma A, Ross MA and Collins AR (1996 Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes. Cancer Res 56: 1291–1295
Glynn SA and Albanes D (1994 Folate and cancer: a review of the literature. Nutr Cancer 22: 101–119
Goulian M, Bleile B and Tseng BY (1980 The effect of methotrexate on levels of dUTP in animal cells. J Biol Chem 255: 10630–10637
Grant G, Dorward PM and Pusztai A (1993 Pancreatic enlargement is evident in rats fed diets containing raw soyabean (Glycine max) or cowpea (Vigna unguiculata) for 800 days but not in those given diets based on kidney bean (Phaseolus vulgaris) or lupinseed (Lupinus angustifolius). J Nutr 123: 2207–2215
James SJ, Miller BJ, Basknakian AG, Pogribny IP, Pogribny M and Muskhelishvili L (1997 Apoptosis and proliferation under conditions of deoxynucleotide pool imbalance in liver of folate/methyl deficient rats. Carcinogenesis 18: 287–293
Kim Y-I (1999 Folate and carcinogenesis: evidence, mechanisms and implications. J Nutr Biochem 10: 6–88
Koury MJ and Horne DW (1994 Apoptosis mediates and thymidine prevents erythroblast destruction in folate deficiency anaemia. P.N.A.S 91: 4067–4071
Koury MJ, Horne DW, Blount BC and Ames BN (1995 Erythroblasts undergoing apoptosis due to folate deficiency misincorporate uracil into DNA and accumulate the tumour suppressor protein p53. Blood 86: 2324
Lowry OH, Rosenburgh NJ, Farr AL and Randall RJ (1951 Protein measurement with folin phenol reagent. J Biol Chem 193: 265–270
Melnyk S, Pogribny M, Miller BJ, Basnakian AG, Pogribny IP and James SJ (1999 Uracil misincorporation, DNA strand breaks and gene amplification are associated with tumorigenic cell transformation in folate deficient/repleted Chinese hamster ovary cells. Cancer Letts 146: 35–44
Nakae D, Kotake Y, Kishida H, Hensley KL, Denda A, Kobayashi Y, Kitayama W, Tsujiuchi T, Sang H, Stewart CA, Tabatbaie T, Floyd RA and Konoshi Y (1998 Inhibition by phenyl N-tert-butyl nitrone of early phase carcinogenesis in the livers of rats fed a choline-deficient, L-amino acid-defined diet. Cancer Res 58: 4548–4551
Pogribny IP, Basnakian AG, Miller BJ, Lopatina NG, Poirier LA and James SJ (1995 Breaks in genomic DNA and within the p53 gene are associated with hypomethylation in livers of folate/methyl deficient rats. Cancer Res 55: 1894–1901
Pogribny IP, Muskheloshvili L, Miller BJ and James SJ (1997 Presence and consequence of uracil in preneoplastic DNA from folate/methyl-deficient rats. Carcinogenesis 18: 2071–2076
Reidy JA (1987 Folate- and deoxyuridine-sensitive chromatid breakage may result from DNA repair during G2. Mutat Res 192: 217–219
Reidy JA (1988 Role of deoxyuridine incorporation and DNA repair in the expression of human chromosomal fragile sites. Mutat Res 200: 215–220
Stolzenberg-Solomon RZ, Albanes D, Nieto FJ, Hartman TJ, Tangrea JA, Rautalahi M, Selhub J, Virtamo J and Taylor PR (1999 Pancreatic cancer risk and nutrition-related methyl group availability indicators in male smokers. J Natl Cancer Inst 91: 535–541
Verhoff P, Stampfer MJ, Buring JE, Gaziano JM, Allen RH, Stabler SP, Reynolds RD, Kok FJ, Hennekens CH and Willet W (1995 Homocysteine metabolism and risk of myocardial infarction: relation with vitamins B6, B12and folate. Am J Epidemiol 143: 845–859
Wickramasinghe SN and Fida S (1993 Misincorporation of uracil into the DNA of folate- and B12-deficient HL60 cells. Eur J Haematol 50: 127–132
Wickramasinghe SN and Fida S (1994 Bone marrow cells from vitamin B12- and folate-deficient patients misincorporate uracil into DNA. Blood 83: 1656–1661
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Duthie, S., Grant, G. & Narayanan, S. Increased uracil misincorporation in lymphocytes from folate-deficient rats. Br J Cancer 83, 1532–1537 (2000). https://doi.org/10.1054/bjoc.2000.1481
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1054/bjoc.2000.1481
Keywords
This article is cited by
-
One-Carbon Metabolism and Colorectal Cancer: Potential Mechanisms of Chemoprevention
Current Pharmacology Reports (2015)
-
A folate receptor alpha double-mutated haplotype 1816delC–1841A is distributed throughout Eurasia and associated with lower erythrocyte folate levels
Molecular Biology Reports (2012)
-
Folate and cancer: how DNA damage, repair and methylation impact on colon carcinogenesis
Journal of Inherited Metabolic Disease (2011)
-
Mitochondrial DNA deletions of blood lymphocytes as genetic markers of low folate-related mitochondrial genotoxicity in peripheral tissues
European Journal of Nutrition (2009)
-
Sensitivity of markers of DNA stability and DNA repair activity to folate supplementation in healthy volunteers
British Journal of Cancer (2006)