Allele and Genotype Frequencies of the Polymorphic Methylenetetrahydrofolate Reductase and Colorectal Cancer among Jordanian Population

  • Yousef, Al-Motassem (Department of Biopharmaceutics and Clinical Pharmacy, Faculty of Pharmacy, The University of Jordan) ;
  • Shomaf, Maha (Department of Pathology, Department of Biochemistry, Faculty of Medicine, The University of Jordan) ;
  • Berger, Sondra (Department of Pharmaceutical and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina) ;
  • Ababneh, Nidaa (Molecular Biology Research Lab, Department of Biochemistry, Faculty of Medicine, The University of Jordan) ;
  • Bobali, Yahya (Molecular Biology Research Lab, Department of Biochemistry, Faculty of Medicine, The University of Jordan) ;
  • Ali, Dema (Molecular Biology Research Lab, Department of Biochemistry, Faculty of Medicine, The University of Jordan) ;
  • Al-Hasan, Sara (Molecular Biology Research Lab, Department of Biochemistry, Faculty of Medicine, The University of Jordan) ;
  • Diab, Ola (Department of Biopharmaceutics and Clinical Pharmacy, Faculty of Pharmacy, The University of Jordan) ;
  • Ismail, Said (Molecular Biology Research Lab, Department of Biochemistry, Faculty of Medicine, The University of Jordan)
  • Published : 2013.08.30


Background: Methylenetetrahydrofolate reductase (MTHFR) is involved in DNA synthesis and repair. We here aimed to investigate two common polymorphisms, C677T and A1298C, with genotype and haplotype frequencies in colorectal cancer (CRC) cases among Jordanian. Materials and Methods: 131 CRC cases were studied for MTHFR C677T and A1298C polymorphisms, compared to 117 controls taken from the general population, employing the PCR-RFLP technique. Results: We found the frequency of the three different genotypes of MTHFR C677T among Jordanians to be CC: 61.7%, CT: 35.2%, and TT 3.1% among CRC cases and 50.9%, 38.8% and 10.3% among controls. Carriers of the TT genotype were less likely to have CRC (OR=0.25; 95%CI: 0.076-0.811; p=0.021) as compared to those with the CC genotype. Genotype analysis of MTHFR A12987C revealed AA: 38.9%, AC: 45%, and CC 16% among CRC cases and 37.4%, 50.4% and 12.2% among controls. There was no significant association between genetic polymorphism at this site and CRC. Haplotype analysis of MTHFR polymorphism at the two loci showed differential distribution of the TA haplotype (677T-1298A) between cases and controls. The TA haplotype was associated with a decreased risk for colorectal cancer (OR=0.6; 95% CI: 0.4-0.9, p=0.03). Conclusions: The genetic polymorphism of MTHFR at 677 and the TA haplotype may modulate the risk for CRC development among the Jordanian population. Our findings may reflect an importance of genes involved in folate metabolism in cancer risk.


Genetic polymorphism;colorectal cancer;methylene tetrahydrofolate;genomic DNA


  1. Arafa MA, Waly MI, Jriesat S, et al (2011). Dietary and lifestyle characteristics of colorectal cancer in Jordan: a case-control study. Asian Pac J Cancer Prev, 12, 1931-6.
  2. Bagley PJ, Selhub J (1998). A common mutation in the methylenetetrahydrofolate reductase gene is associated with an accumulation of formylated tetrahydrofolates in red blood cells. Proc Natl Acad Sci USA, 95, 13217-20.
  3. Bailey LB (2003). Folate, methyl-related nutrients, alcohol, and the MTHFR 677C-->T polymorphism affect cancer risk: intake recommendations. J Nutr, 133, 3748-53.
  4. Brockton NT (2006). Localized depletion: the key to colorectal cancer risk mediated by MTHFR genotype and folate? Cancer Causes Control, 17, 1005-16.
  5. Brooker R, Ed. (2005). Genetics, Analysis and Principles. United State of America, McGraw-Hill.
  6. Caccamo D, Condello S, Gorgone G, et al (2004). Screening for C677T and A1298C MTHFR polymorphisms in patients with epilepsy and risk of hyperhomocysteinemia. Neuromolecular Med, 6, 117-26.
  7. Center MM, Jemal A, Ward E (2009). International trends in colorectal cancer incidence rates. Cancer Epidemiol Biomarkers Prev, 18, 1688-94.
  8. Central Intelligence Agency. (2012). The World Factbook.Retrieved Feb 18th, 2012, from
  9. Chandy S, Sadananda Adiga MN, Ramachandra N, et al (2010). Association of methylenetetrahydrofolate reductase gene polymorphisms and colorectal cancer in India. Indian J Med Res, 131, 659-64.
  10. Chango A, Boisson F, Barbe F, et al (2000). The effect of 677C-->T and 1298A-->C mutations on plasma homocysteine and 5,10-methylenetetrahydrofolate reductase activity in healthy subjects. Br J Nutr, 83, 593-6.
  11. Chen J, Giovannucci E, Hankinson SE, et al (1998). A prospective study of methylenetetrahydrofolate reductase and methionine synthase gene polymorphisms, and risk of colorectal adenoma. Carcinogenesis, 19, 2129-32.
  12. Chen J, Giovannucci E, Kelsey K, et al (1996). A methylenetetrahydrofolate reductase polymorphism and the risk of colorectal cancer. Cancer Res, 56, 4862-4.
  13. Chen J, Ma J, Stampfer MJ, et al (2002). Linkage disequilibrium between the 677C>T and 1298A>C polymorphisms in human methylenetetrahydrofolate reductase gene and their contributions to risk of colorectal cancer. Pharmacogenetics, 12, 339-42.
  14. Cicek MS, Nock NL, Li L, et al (2004). Relationship between methylenetetrahydrofolate reductase C677T and A1298C genotypes and haplotypes and prostate cancer risk and aggressiveness. Cancer Epidemiol Biomarkers Prev, 13, 1331-6.
  15. Curtin K, Bigler J, Slattery ML, et al (2004). MTHFR C677T and A1298C polymorphisms: diet, estrogen, and risk of colon cancer. Cancer Epidemiol Biomarkers Prev, 13, 285-92.
  16. El Awady MK, Karim AM, Hanna LS, et al (2009). Methylenetetrahydrofolate reductase gene polymorphisms and the risk of colorectal carcinoma in a sample of Egyptian individuals. Cancer Biomark, 5, 233-40.
  17. Ferlay J, Shin HR, Bray F, et al (2008). GLOBOCAN 2008 v1.2, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10 [Internet]. Retrieved June 2012, 2012, from
  18. Ferlay J, Shin HR, Bray F, et al (2010). GLOBOCAN 2008 v1.2, cancer incidence and mortality worldwide: Iarc cancerbase no. 10 [internet]. Retrieved March, 2013, from
  19. Friedman G, Goldschmidt N, Friedlander Y, et al (1999). A common mutation A1298C in human methylenetetrahydrofolate reductase gene: association with plasma total homocysteine and folate concentrations. J Nutr, 129, 1656-61.
  20. Frosst P, Blom HJ, Milos R, et al (1995). A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet, 10, 111-3.
  21. Goyette P, Pai A, Milos R, et al (1998). Gene structure of human and mouse methylenetetrahydrofolate reductase (MTHFR). Mamm Genome, 9, 652-6.
  22. Graph Pad Software Inc. Retrieved 20th February, 2011, from
  23. Guimaraes JL, Ayrizono Mde L, Coy CS, Lima CS (2011). Gene polymorphisms involved in folate and methionine metabolism and increased risk of sporadic colorectal adenocarcinoma. Tumour Biol, 32, 853-61.
  24. Houlston RS, Tomlinson IP (2001). Polymorphisms and colorectal tumor risk. Gastroenterology, 121, 282-301.
  25. Keku T, Millikan R, Worley K, et al (2002). 5, 10-Methylenetetrahydrofolate reductase codon 677 and 1298 polymorphisms and colon cancer in African Americans and whites. Cancer Epidemiol Biomarkers Prev, 11, 1611-21.
  26. Klerk M, Verhoef P, Clarke R, et al (2002). MTHFR 677C-->T polymorphism and risk of coronary heart disease: a meta-analysis. JAMA, 288, 2023-31.
  27. Kono S, Chen K (2005). Genetic polymorphisms of methylenetetrahydrofolate reductase and colorectal cancer and adenoma. Cancer Sci, 96, 535-42.
  28. Le Marchand L, Donlon T, Hankin JH, et al (2002). B-vitamin intake, metabolic genes, and colorectal cancer risk (United States). Cancer Causes Control, 13, 239-48.
  29. Levine AJ, Siegmund KD, Ervin CM, et al (2000). The methylenetetrahydrofolate reductase 677C-->T polymorphism and distal colorectal adenoma risk. Cancer Epidemiol Biomarkers Prev, 9, 657-63.
  30. Li H, Xu WL, Shen HL, et al (2011). Methylenetetrahydrofolate reductase genotypes and haplotypes associated with susceptibility to colorectal cancer in an eastern Chinese Han population. Genet Mol Res, 10, 3738-46.
  31. Lievers KJ, Boers GH, Verhoef P, et al (2001). A second common variant in the methylenetetrahydrofolate reductase (MTHFR) gene and its relationship to MTHFR enzyme activity, homocysteine, and cardiovascular disease risk. J Mol Med (Berl), 79, 522-8.
  32. Lowi MR (1995). Water and power: the politics of a scarce resource in the Jordan River basin. Cambridge, Cambridge University Press.
  33. Lucock M (2000). Folic acid: nutritional biochemistry, molecular biology, and role in disease processes. Mol Genet Metab, 71, 121-38.
  34. Lucock M (2004). Is folic acid the ultimate functional food component for disease prevention? BMJ, 328, 211-4.
  35. Ma J, Stampfer MJ, Giovannucci E, et al (1997). Methylenetetrahydrofolate reductase polymorphism, dietary interactions, and risk of colorectal cancer. Cancer Res, 57, 1098-102.
  36. Marugame T, Tsuji E, Kiyohara C, et al (2003). Relation of plasma folate and methylenetetrahydrofolate reductase C677T polymorphism to colorectal adenomas. Int J Epidemiol, 32, 64-6.
  37. Meisel C, Cascorbi I, Gerloff T, et al (2001). Identification of six methylenetetrahydrofolate reductase (MTHFR) genotypes resulting from common polymorphisms: impact on plasma homocysteine levels and development of coronary artery disease. Atherosclerosis, 154, 651-8.
  38. Ogino S, Wilson RB (2003). Genotype and haplotype distributions of MTHFR677C>T and 1298A>C single nucleotide polymorphisms: a meta-analysis. J Hum Genet, 48, 1-7.
  39. Pardini B, Kumar R, Naccarati A, et al (2011). MTHFR and MTRR genotype and haplotype analysis and colorectal cancer susceptibility in a case-control study from the Czech Republic. Mutat Res, 721, 74-80.
  40. Park KS, Mok JW, Kim JC (1999). The 677C > T mutation in 5,10-methylenetetrahydrofolate reductase and colorectal cancer risk. Genet Test, 3, 233-6.
  41. Parkin DM, Pisani P, Ferlay J (1999). Estimates of the worldwide incidence of 25 major cancers in 1990. Int J Cancer, 80, 827-41.<827::AID-IJC6>3.0.CO;2-P
  42. Sachse C, Smith G, Wilkie MJ, et al (2002). A pharmacogenetic study to investigate the role of dietary carcinogens in the etiology of colorectal cancer. Carcinogenesis, 23, 1839-49.
  43. Shannon B, Gnanasampanthan S, Beilby J, Iacopetta B (2002). A polymorphism in the methylenetetrahydrofolate reductase gene predisposes to colorectal cancers with microsatellite instability. Gut, 50, 520-4.
  44. Sharp L, Little J (2004). Polymorphisms in genes involved in folate metabolism and colorectal neoplasia: a HuGE review. Am J Epidemiol, 159, 423-43.
  45. Shen H, Spitz MR, Wang LE, et al (2001). Polymorphisms of methylene-tetrahydrofolate reductase and risk of lung cancer: a case-control study. Cancer Epidemiol Biomarkers Prev, 10, 397-401.
  46. Shi M, Caprau D, Romitti P, et al (2003). Genotype frequencies and linkage disequilibrium in the CEPH human diversity panel for variants in folate pathway genes MTHFR, MTHFD, MTRR, RFC1, and GCP2. Birth Defects Res A Clin Mol Teratol, 67, 545-9.
  47. Slattery ML, Potter JD, Samowitz W, et al (1999). Methylenetetrahydrofolate reductase, diet, and risk of colon cancer. Cancer Epidemiol Biomarkers Prev, 8, 513-8.
  48. Stegmann K, Ziegler A, Ngo ET, et al (1999). Linkage disequilibrium of MTHFR genotypes 677C/T-1298A/C in the German population and association studies in probands with neural tube defects(NTD). Am J Med Genet, 87, 23-9.<23::AID-AJMG5>3.0.CO;2-E
  49. Taioli E, Garza MA, Ahn YO, et al (2009). Meta- and pooled analyses of the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and colorectal cancer: a HuGE-GSEC review. Am J Epidemiol, 170, 1207-21.
  50. Terrazzino S, Agostini M, Pucciarelli S, et al (2006). A haplotype of the methylenetetrahydrofolate reductase gene predicts poor tumor response in rectal cancer patients receiving preoperative chemoradiation. Pharmacogenet Genomics, 16, 817-24.
  51. The Royal Hashemite Court. "Keys to the Kingdom-The People of Jordan." Retrieved Feb 18th, 2012, from
  52. van der Put NM, Gabreels F, Stevens EM, et al (1998). A second common mutation in the methylenetetrahydrofolate reductase gene: an additional risk factor for neural-tube defects? Am J Hum Genet, 62, 1044-51.
  53. Weisberg I, Tran P, Christensen B, et al (1998). A second genetic polymorphism in methylenetetrahydrofolate reductase (MTHFR) associated with decreased enzyme activity. Mol Genet Metab, 64, 169-72.
  54. Yi P, Pogribny I, Jill James S (2002). Multiplex PCR for simultaneous detection of 677 C-->T and 1298 A-->C polymorphisms in methylenetetrahydrofolate reductase gene for population studies of cancer risk. Cancer Lett, 181, 209.
  55. Yin G, Kono S, Toyomura K, et al (2004). Methylenetetrahydrofolate reductase C677T and A1298C polymorphisms and colorectal cancer: the Fukuoka Colorectal Cancer Study. Cancer Sci, 95, 908-13.
  56. Zhou D, Mei Q, Luo H, et al (2012). The polymorphisms in methylenetetrahydrofolate reductase, methionine synthase, methionine synthase reductase, and the risk of colorectal cancer. Int J Biol Sci, 8, 819-30.

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