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. 2000 Nov 22;83(11):1532–1537. doi: 10.1054/bjoc.2000.1481

Increased uracil misincorporation in lymphocytes from folate-deficient rats

S J Duthie 1, G Grant 1, S Narayanan 1
PMCID: PMC2363426  PMID: 11076664

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

Keywords: folic acid deficiency, methyl-donor deficiency, rat lymphocytes, misincorporated uracil, oxidized DNA bases

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Selected References

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  1. Albright C. D., Liu R., Mar M. H., Shin O. H., Vrablic A. S., Salganik R. I., Zeisel S. H. Diet, apoptosis, and carcinogenesis. Adv Exp Med Biol. 1997;422:97–107. doi: 10.1007/978-1-4757-2670-1_8. [DOI] [PubMed] [Google Scholar]
  2. Asahara H., Wistort P. M., Bank J. F., Bakerian R. H., Cunningham R. P. Purification and characterization of Escherichia coli endonuclease III from the cloned nth gene. Biochemistry. 1989 May 16;28(10):4444–4449. doi: 10.1021/bi00436a048. [DOI] [PubMed] [Google Scholar]
  3. Blount B. C., Ames B. N. Analysis of uracil in DNA by gas chromatography-mass spectrometry. Anal Biochem. 1994 Jun;219(2):195–200. doi: 10.1006/abio.1994.1257. [DOI] [PubMed] [Google Scholar]
  4. Blount B. C., Mack M. M., Wehr C. M., MacGregor J. T., Hiatt R. A., Wang G., Wickramasinghe S. N., Everson R. B., Ames B. N. Folate deficiency causes uracil misincorporation into human DNA and chromosome breakage: implications for cancer and neuronal damage. Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3290–3295. doi: 10.1073/pnas.94.7.3290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Czeizel A. E. Prevention of congenital abnormalities by periconceptional multivitamin supplementation. BMJ. 1993 Jun 19;306(6893):1645–1648. doi: 10.1136/bmj.306.6893.1645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duthie S. J. Folic acid deficiency and cancer: mechanisms of DNA instability. Br Med Bull. 1999;55(3):578–592. doi: 10.1258/0007142991902646. [DOI] [PubMed] [Google Scholar]
  7. Duthie S. J., Hawdon A. DNA instability (strand breakage, uracil misincorporation, and defective repair) is increased by folic acid depletion in human lymphocytes in vitro. FASEB J. 1998 Nov;12(14):1491–1497. [PubMed] [Google Scholar]
  8. Duthie S. J., Ma A., Ross M. A., Collins A. R. Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes. Cancer Res. 1996 Mar 15;56(6):1291–1295. [PubMed] [Google Scholar]
  9. Glynn S. A., Albanes D. Folate and cancer: a review of the literature. Nutr Cancer. 1994;22(2):101–119. doi: 10.1080/01635589409514336. [DOI] [PubMed] [Google Scholar]
  10. Goulian M., Bleile B., Tseng B. Y. The effect of methotrexate on levels of dUTP in animal cells. J Biol Chem. 1980 Nov 25;255(22):10630–10637. [PubMed] [Google Scholar]
  11. Grant G., Dorward P. M., Pusztai A. Pancreatic enlargement is evident in rats fed diets containing raw soybeans (Glycine max) or cowpeas (Vigna unguiculata) for 800 days but not in those fed diets based on kidney beans (Phaseolus vulgaris) or lupinseed (Lupinus angustifolius). J Nutr. 1993 Dec;123(12):2207–2215. doi: 10.1093/jn/123.12.2207. [DOI] [PubMed] [Google Scholar]
  12. James S. J., Miller B. J., Basnakian A. G., Pogribny I. P., Pogribna M., Muskhelishvili L. Apoptosis and proliferation under conditions of deoxynucleotide pool imbalance in liver of folate/methyl deficient rats. Carcinogenesis. 1997 Feb;18(2):287–293. doi: 10.1093/carcin/18.2.287. [DOI] [PubMed] [Google Scholar]
  13. Kim Y. I. Folate and carcinogenesis: evidence, mechanisms, and implications. J Nutr Biochem. 1999 Feb;10(2):66–88. doi: 10.1016/s0955-2863(98)00074-6. [DOI] [PubMed] [Google Scholar]
  14. Koury M. J., Horne D. W. Apoptosis mediates and thymidine prevents erythroblast destruction in folate deficiency anemia. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):4067–4071. doi: 10.1073/pnas.91.9.4067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Melnyk S., Pogribna M., Miller B. J., Basnakian A. G., Pogribny I. P., James S. J. Uracil misincorporation, DNA strand breaks, and gene amplification are associated with tumorigenic cell transformation in folate deficient/repleted Chinese hamster ovary cells. Cancer Lett. 1999 Nov 1;146(1):35–44. doi: 10.1016/s0304-3835(99)00213-x. [DOI] [PubMed] [Google Scholar]
  17. Nakae D., Kotake Y., Kishida H., Hensley K. L., Denda A., Kobayashi Y., Kitayama W., Tsujiuchi T., Sang H., Stewart C. A. 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. 1998 Oct 15;58(20):4548–4551. [PubMed] [Google Scholar]
  18. Pogribny I. P., Basnakian A. G., Miller B. J., Lopatina N. G., Poirier L. A., James S. J. Breaks in genomic DNA and within the p53 gene are associated with hypomethylation in livers of folate/methyl-deficient rats. Cancer Res. 1995 May 1;55(9):1894–1901. [PubMed] [Google Scholar]
  19. Pogribny I. P., Muskhelishvili L., Miller B. J., James S. J. Presence and consequence of uracil in preneoplastic DNA from folate/methyl-deficient rats. Carcinogenesis. 1997 Nov;18(11):2071–2076. doi: 10.1093/carcin/18.11.2071. [DOI] [PubMed] [Google Scholar]
  20. Reidy J. A. Folate- and deoxyuridine-sensitive chromatid breakage may result from DNA repair during G2. Mutat Res. 1987 Nov;192(3):217–219. doi: 10.1016/0165-7992(87)90059-5. [DOI] [PubMed] [Google Scholar]
  21. Reidy J. A. Role of deoxyuridine incorporation and DNA repair in the expression of human chromosomal fragile sites. Mutat Res. 1988 Jul-Aug;200(1-2):215–220. doi: 10.1016/0027-5107(88)90085-1. [DOI] [PubMed] [Google Scholar]
  22. Stolzenberg-Solomon R. Z., Albanes D., Nieto F. J., Hartman T. J., Tangrea J. A., Rautalahti M., Sehlub J., Virtamo J., Taylor P. R. Pancreatic cancer risk and nutrition-related methyl-group availability indicators in male smokers. J Natl Cancer Inst. 1999 Mar 17;91(6):535–541. doi: 10.1093/jnci/91.6.535. [DOI] [PubMed] [Google Scholar]
  23. Verhoef P., Stampfer M. J., Buring J. E., Gaziano J. M., Allen R. H., Stabler S. P., Reynolds R. D., Kok F. J., Hennekens C. H., Willett W. C. Homocysteine metabolism and risk of myocardial infarction: relation with vitamins B6, B12, and folate. Am J Epidemiol. 1996 May 1;143(9):845–859. doi: 10.1093/oxfordjournals.aje.a008828. [DOI] [PubMed] [Google Scholar]
  24. Wickramasinghe S. N., Fida S. Bone marrow cells from vitamin B12- and folate-deficient patients misincorporate uracil into DNA. Blood. 1994 Mar 15;83(6):1656–1661. [PubMed] [Google Scholar]
  25. Wickramasinghe S. N., Fida S. Misincorporation of uracil into the DNA of folate- and B12-deficient HL60 cells. Eur J Haematol. 1993 Mar;50(3):127–132. doi: 10.1111/j.1600-0609.1993.tb00080.x. [DOI] [PubMed] [Google Scholar]
  26. de Bree A., van Dusseldorp M., Brouwer I. A., van het Hof K. H., Steegers-Theunissen R. P. Folate intake in Europe: recommended, actual and desired intake. Eur J Clin Nutr. 1997 Oct;51(10):643–660. doi: 10.1038/sj.ejcn.1600467. [DOI] [PubMed] [Google Scholar]

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