MTHFR C677T Polymorphism and Recurrent Early Pregnancy Loss Risk in North Indian Population


Recurrent early pregnancy loss (REPL) is a multifactorial disorder as both genetic and environmental factors contribute to the development of disease. Folate metabolism is an important mechanism to ensure proper fetal growth. Hyperhomocysteinemia leads to a number of disorders and REPL is one of them. In a case–control study DNA from 106 cases with the history of 3 or more REPL and 140 healthy fertile controls with successful pregnancy outcomes were genotyped for C677T single-nucleotide polymorphism (SNP) of the MTHFR (methylenetetrahydrofolate reductase) gene through polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP), which was further confirmed by sequencing. Allele frequencies of REPL cases were compared with healthy controls and a statistically significant association was found between REPL and the mutant T allele (χ2 = 8.786, odds ratio [OR] = 2.20, 95% confidence interval [CI] = 1.323-3.9658, P = .003). The genotype frequencies of SNP C677T also differ significantly between these 2 groups (χ2 = 8.237, P = .016). The OR for heterozygous CT in the REPL versus controls is 1.9591 (95% CI = 1.0285-3.7318, P = .04). The OR for TT homozygous is 6.3009 (95% CI = 1.2065, P = .02). Combined odds ratio of CT and TT against the control has been calculated as 2.2194 (95% CI = 1.2029-4.0952, P = .02) which is also significant. Thus the present study clearly indicates that homozygosity and heterozygosity for the MTHFR C677T polymorphism confer a 6.3009- and 1.9591-fold increased risk of idiopathic REPL, respectively.

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  1. 1.

    Laird SM, Tuckerman EM, Cork BA, Linjawi S, Blakemore AIF, Li TC. A review of immune cells and molecules in women with recurrent miscarriage. Hum Reprod Update. 2003;9(2):163–174.

  2. 2.

    Extalo N. Early human nutrition. Eur J Obstet Gyn R B. 1995;61: 3–6.

  3. 3.

    Nelen WL, Bulten J, Steegers EA, Blom HJ, Hanselaar AG, Eskes TK. Maternal homocysteine and chorionic vascularization in recurrent early pregnancy loss. Hum Reprod. 2000;15(4):954–960.

  4. 4.

    Choi HK, Choi BC, Lee SH, Kim JW, Cha KY, Baek KH. Expression of angiogenesis- and apoptosis-related genes in chorionic villi derived from recurrent pregnancy patients. Mol Reprod Dev. 2003;66(1):24–31.

  5. 5.

    Bailey L, Gregory J. Polymorphisms of methylenetetrahydrofolate reductase and other enzymes: metabolic significance, risks and impact on folate requirement. J Nutr. 1999;129(5):919–922.

  6. 6.

    Wenstrom K, Johanning G, Owen J, et al. Amniotic fluid homocysteine levels, 5,10-methylenetetrahydrafolate reductase genotypes, and neural tube closure sites. Am J Med Genet. 2000; 90(1):6–11.

  7. 7.

    Wang J, Trudinger B, Duarte N, Wilcken D, Wang X. Elevated circulating homocyst(e)ine levels in placental vascular disease and associated pre-eclampsia. BJOG. 2000;107(7):935–938.

  8. 8.

    Ray J, Laskin C. Folic acid and homocyst(e)ine metabolic defects and the risk of placental abruption, pre-eclampsia and spontaneous pregnancy loss: a systematic review. Placenta. 1999;20(7):519–529.

  9. 9.

    Sohda S, Arinami T, Hamada H, Yamada N, Hamaguchi H, Kubo T. Methylenetetrahydrofolate reductase polymorphism and preecla psia. J Med Genet. 1997;34(6):525–526.

  10. 10.

    McCully K. Chemical pathology of homocysteine. II. Carcinogenesis and homocysteine thiolactone metabolism. Ann Clin Lab Sci. 1994;24(1):27–59.

  11. 11.

    Wouters MG, Boers GH, Blom HJ, et al. Hyperhomocysteinemia: a risk factor in women with unexplained recurrent early pregnancy loss. Fertil Steril. 1993;60(5):820–825.

  12. 12.

    Cai D, Ning L, Pan C, et al. Association of polymorphisms in folate metabolic genes and prostate Cancer risk: a case–control study in a Chinese population. J Genet. 2010;89(2):263–267.

  13. 13.

    Obwegeser R, Sinzinger HM. Homocysteine—a pathophysiological cornerstone in obstetrical and gynaecological disorders? Hum Reprod Update. 1999;5(1):64–72.

  14. 14.

    Frosst P, Blom H, Milos R, et al. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet. 1995;10(1):111–113.

  15. 15.

    Weisberg I, Tran P, Christensen B, Sibani S, Rozen R. A second genetic polymorphism in methylenetetrahydrofolate reductase (MTHFR) associated with decreased enzyme activity. Mol.Genet Metab. 1998;64(3):169–172.

  16. 16.

    Mostowska A, Hozyasz KK, Wojcicki P, Dziegelewska M, Jagodzinski PP. Associations of folate and choline metabolism gene polymorphisms with orofacial clefts. J Med Genet. 2009; 47(12):809–815. DOI: 10.1136/jmg.2009.070029.

  17. 17.

    Faul F, Erdfelder E, Buchner A, Lang AG. Statistical power analyses using G*Power 3.1: tests for correlation and regression analyses. Behav Res Methods. 2009;41(4):1149–1160.

  18. 18.

    Faul F, Erdfelder E, Lang AG, Buchner A. G*Power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods. 2007;39(2):175–191.

  19. 19.

    Fryer RH, Wilson BD, Gubler DB, Fitzgerald LA, Rodgers GM. Homocysteine, a risk factor for premature vascular disease and thrombosis, induces tissue factor activity in endothelial cells. Arterioscler Thromb. 1993;13(9):1327–1333.

  20. 20.

    Poddar R, Sivasubramanian N, DiBello PM, Robinson K, Jacobsen DW. Homocysteine induces expression and secretion of monocyte chemoattractant protein-1 and interleukin-8 in human aortic endothelial cells: implications for vascular disease. Circulation. 2001;103(22):2717–2723.

  21. 21.

    Hefler L, Jirecek S, Heim K, et al. Genetic polymorphisms associated with thrombophilia and vascular disease in women with unexplained late intrauterine fetal death: a multicenter study. J Soc Gynecol Investig. 2004;11(1):42–44.

  22. 22.

    Rey E, Kahn SR, David M, Shrier I. Thrombophilic disorders and fetal loss: a meta-analysis. Lancet. 2003;361(9361):901–908.

  23. 23.

    Lin J, August P. Genetic thrombophilias and preeclampsia: a meta-analysis. Obstet Gynecol. 2005;105(1):182–192.

  24. 24.

    Tempfer C, Schneeberger C, Riener EK, Hefler LA, Keck C. Polymorphisms for risk assessment of pregnancy-associated disorders: review of the literature and meta-analysis. Geburtsh Frauenheilk. 2004;64(10):1080–1089.

  25. 25.

    Van der Molen EF, Arends GE, Nelen WL, et al. A common mutation in the 5,10-methylenetetrahydrofolate reductase gene as a new risk factor for placental vasculopathy. Am J Obstet Gynecol. 2000;182(5):1258–1263.

  26. 26.

    Unfried G, Griesmacher A, Weismuller W, Nagele F, Huber JC, Tempfer CB. The C677T polymorphism of the methylenetetrahydrofolate reductase gene and idiopathic recurrent miscarriage. Obstet Gynecol. 2002;99(4):614–619.

  27. 27.

    Nelen WL, Steegers EA, Eskes TK, Blom HJ. Genetic risk factor for unexplained recurrent early pregnancy loss. Lancet. 1997; 350(9081):861.

  28. 28.

    Quere I, Bellet H, Hoffet M, Janbon C, Mares P, Gris JC. A woman with five consecutive fetal deaths: case report and retrospective analysis of hyperhomocysteinaemia prevalence in 100 consecutive women with recurrent miscarriages. Fertil Steril. 1998;69(1):152–154.

  29. 29.

    Sarig G, Younis JS, Hoffman R, Lanir N, Blumenfeld Z, Brenner B. Thrombophilia is common in women with idiopathic pregnancy loss and is associated with late pregnancy wastage. Fertil Steril. 2002;77(2):342–347.

  30. 30.

    Foka ZJ, Lambropoulos AF, Saravelos H, et al. Factor V Leiden and prothrombin G20210A mutations, but not methylenetetrahydrofolate reductase C677T, are associated with recurrent miscarriages. Hum Reprod. 2000;15(2):458–462.

  31. 31.

    Makino A, Nakanishi T, Sugiura-Ogasawara M, Ozaki Y, Suzumori N, Suzumori K. No association of C677T methylenetetrahydrofolate reductase and an endothelial nitric oxide synthase polymorphism with recurrent pregnancy loss. Am J Reprod Immunol. 2004;52(1):60–66.

  32. 32.

    Kutteh WH, Park VM, Deitcher SR. Hypercoagulable state mutation analysis in white patients with early first-trimester recurrent pregnancy loss. Fertil Steril. 1999;71(6):1048–1053.

  33. 33.

    Carp H, Salomon O, Seidman D, Dardik R, Rosenberg N, Inbal A. Prevalence of genetic markers for thrombophilia in recurrent pregnancy loss. Hum Reprod. 2002;17(6):1633–1637.

  34. 34.

    Biswas A, Choudhry P, Mittal A, et al. Recurrent abortions in Asian Indians: no role of factor V Leiden Hong Kong/Cambridge mutation and MTHFR polymorphism. Clin Appl Thromb Hemost. 2008;4(1):102–104.

  35. 35.

    Mukhopadhyay R, Saraswathy KN, Ghosh PK. MTHFR C677T and factor V Leiden in recurrent pregnancy loss: a study among an endogamous group in north India. Genet Test Mol Biomarkers. 2009;13(6):861–865.

  36. 36.

    Kumar KSD, Govindaiah V, Naushad SE, Devi RR, Jyothy A. Plasma homocysteine levels correlated to interactions between folate status and methylene tetrahydrofolate reductase gene mutation in women with unexplained recurrent pregnancy loss. J Obstet Gynecol. 2003;23(1):55–58.

  37. 37.

    Lakshmi AV, Maniprabha C, Krishna TP. Plasma homocysteine level in relation to folate and vitamin B6 status in apparently normal men. Asia Pac J Clin Nutr. 2001;10(3):194–196.

  38. 38.

    Refsum M, Yajnik CS, Gadkari M, et al. Hyperhomocysteinaemia and elevated methylmalonic acid indicate a high prevalence of cobalamin deficiency in Asian Indians. Am J Clin Nutr. 2001; 74(2):233–241.

  39. 39.

    Singh K, Singh SK, Sah R, Singh I, Raman R. Mutation C677T in the methylenetetrahydrofolate reductase gene is associated with male infertility in an Indian population. Int J Androl. 2005; 28(2):115–119.

  40. 40.

    Habibzadeh N, Schorah CJ, Smithells RW. The effects of maternal folic acid and vitamin C nutrition in early pregnancy on reproductive performance in the guinea-pig. Br J Nutr. 1986;55(1): 23–35.

  41. 41.

    Tremblay GF, Matte JJ, Dufour JJ, Brisson GJ. Survival rate and development of fetuses during the first 30 days of gestation after folic acid addition to a swine diet. J Anim Sci. 1989;67(3): 724–732.

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Correspondence to Kiran Singh PhD.

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Nair, R.R., Khanna, A. & Singh, K. MTHFR C677T Polymorphism and Recurrent Early Pregnancy Loss Risk in North Indian Population. Reprod. Sci. 19, 210–215 (2012).

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  • recurrent early pregnancy loss
  • C677T SNP
  • hyperhomocysteinemia
  • angiogenesis