한국인의 반복자연유산 환자에서 Thymidylate Synthase Enhancer Region (TSER) 돌연변이형의 혈중 호모시스테인 양과의 관련성

Contribution of Thymidylate Synthase Enhancer Region (TSER) Polymorphism to Total Plasma Homocysteine Levels in Korean Patients with Recurrent Spontaneous Abortion

  • 최윤경 (포천중문 의과대학교 분당 차병원 진단검사의학과) ;
  • 강명서 (포천중문 의과대학교 분당 차병원 진단검사의학과) ;
  • 김남근 (포천중문 의과대학교 분당 차병원 임상의학연구소) ;
  • 김선희 (포천중문 의과대학교 분당 차병원 임상의학연구소) ;
  • 최동희 (포천중문 의과대학교 분당 차병원 차병원 산부인과 및 여성의학연구소) ;
  • 안명옥 (포천중문 의과대학교 분당 차병원 차병원 산부인과 및 여성의학연구소) ;
  • 이수만 (포천중문 의과대학교 분당 차병원 차병원 산부인과 및 여성의학연구소)
  • Choi, Yoon-Kyung (Department of Diagnostics, Bundang CHA General Hospital, College of Medicine, Pochon CHA University) ;
  • Kang, Myung-Seo (Department of Diagnostics, Bundang CHA General Hospital, College of Medicine, Pochon CHA University) ;
  • Kim, Nam-Keun (Institute for Clinical Research, Department of Obstetrics & Gynecology, Bundang CHA General Hospital, College of Medicine, Pochon CHA University) ;
  • Kim, Sun-Hee (Institute for Clinical Research, Department of Obstetrics & Gynecology, Bundang CHA General Hospital, College of Medicine, Pochon CHA University) ;
  • Choi, Dong-Hee (Infertility Medical Center, Bundang CHA General Hospital, College of Medicine, Pochon CHA University) ;
  • An, Myung-Ok (Infertility Medical Center, Bundang CHA General Hospital, College of Medicine, Pochon CHA University) ;
  • Lee, Su-Man (Infertility Medical Center, Bundang CHA General Hospital, College of Medicine, Pochon CHA University)
  • 발행 : 2004.09.30

초록

Objectives: Methylenetetrahydrofolate reductase (MTHFR) mutation are commonly associated with hyperhomocysteinemia, and through their defects in homocysteine metabolism, they have been implicated as a risk factor for recurrent spontaneous abortion. Recent report describe that 28-bp tandem repeat polymorphism in thymidylate synthase enhancer region (TSER) that influence enzyme activity would affect plasma homocysteine level. We have investigated the relationship between TSER genotype and plasma homocysteine level in 54 patients with recurrent spontaneous abortion. Methods: Plasma homocysteine level was measured by fluorescent polarizing immunoassay. MTHFR mutation (C677T and A1298C) was identified by PCR-restriction fragment length polymorphism assay and TSER mutation was analyzed by PCR method. The data were analyzed using the program SAS 8.2 for Windows. Results: Total homocysteine level was significantly higher in MTHFR 677TT genotype ($9.80{\pm}3.87{\mu}mol/L$) than MTHFR 677CC genotype ($8.14{\pm}1.74{\mu}mol/L$) in Korean patients with unexplained recurrent spontaneous abortion (p=0.0143). However, the plasma homocysteine level was not significantly different in the MTHFR 1298AA ($8.42{\pm}2.65{\mu}mol/L$) and 1298CC ($6.09{\pm}0.32{\mu}mol/L$; p=0.2058) and, TSER 2R2R ($8.61{\pm}1.68{\mu}mol/L$) and 3R3R ($8.05{\pm}2.81{\mu}mol/L$; p=0.9319) mutant genotypes, respectively. In this study, we found the combination effects of TSER and MTHFR C677T genotypes. Plasma homocysteine levels were the highest ($11.47{\pm}4.66{\mu}mol/L$) in individuals with TSER 3R3R ($8.05{\pm}2.81{\mu}mol/L$) and MTHFR 677TT ($9.80{\pm}3.87{\mu}mol/L$) genotypes. Individuals with a combination of both TSER 2R2R/2R3R and MTHFR 677CC/CT genotypes ($7.69{\pm}1.77{\mu}mol/L$) had lower plasma homocysteine levels than TSER 2R2R ($8.61{\pm}1.68{\mu}mol/L$) and MTHR 677CC ($8.14{\pm}1.74{\mu}mol/L$) genotypes, respectively. The effect of MTHFR polymorphism in the homocysteine metabolism appears to be stronger than that of TSER polymorphism. Conclusion: Although statistically not significant, we found the elevated level of plasma homocysteine in combined genotypes with TSER and MTHFR (C677T and A1298C) in Korean patients with unexplained habitual abortion. In this study, we reported the possibility that TSER polymorphism is a genetic determinant of plasma homocysteine levels in the Korean patients as well as MTHFR C677T polymorphism. A large prospective study is needed to verify our findings.

키워드

참고문헌

  1. Younis JS, Ohel G, Brenner B, Ben-Ami M. Fami- lial thrombophilia - the scientific rationale for throm- bophylaxis in recurrent pregnancy loss? Hum Re- prod 1997; 12(7): 1389-90
  2. Raziel A, Kormberg Y, Fridler S, Schachter M, Sela BA, Ron-El R. Hypercoagulable thrombothilic de- fects and hyperhomocysteinemia in patients with recurrent pregnancy loss. Am J Reprod lmmunol 2001; 45(2): 65-71 https://doi.org/10.1111/j.8755-8920.2001.450201.x
  3. Yamada H, Kato EH, Kobashi G, Ebina Y, Shimada S, Sakuragi N, et al. Recurrent pregnancy loss: Etio- logy of thrombophilia. Semin Thromb Hemost 2001; 27: 121-9 https://doi.org/10.1055/s-2001-14070
  4. Nelen WLMD, Blom HJ, Streeers EAP, den Heijer M, Eskes TKAB. yperhomocysteinemia and re- current early pregnancy loss: a meta analysis. Fertil Steril 2000; 74(6): 1196-99 https://doi.org/10.1016/S0015-0282(00)01595-8
  5. McCully KS. Chemical pathology of homocysteine. Ann Clin Lab Sci 1994; 24: 27-59
  6. Chango A, Boisson F, B arbe F, Quilliot D, Droesch S, Pfister M, et al. The effect of 677C->T and 1298A->C mutations on plasma homocysteine and 5,10-methylenetetrahydrofolate reductase activity in healthy subjects. Br J Nutr 2000; 83(6): 593-6 https://doi.org/10.1017/S0007114500000751
  7. Loscalzo J. The oxidant stress of hyperhomocyste- nemia. J Clin Invest 1996; 98: 5-7 https://doi.org/10.1172/JCI118776
  8. Fodinger M, Wagner OF, Horl WH, Sunder-Plass- mann G. Recent insights into the molecular genetics of the homocysteine metabolism. Kidney Int 59 Suppl 2001; 78: S238-42
  9. Jakubowski H, Zhang L, Bardeguez A, Aviv A. Homocysteine thiolactone and proteine homocy- stinylation in human endothilial cells: implications for atherosclerosis. Cir Res 2000; 87(1): 45-57 https://doi.org/10.1161/01.RES.87.1.45
  10. Matsuo K, Suzuki R, Hamajima N, Ogura M, Ka- gami Y, Taji H, et al. Association between poly- morphisms of folate and methionine-metabolizing enzymes and susceptibility to maligant lymphoma. Blood 2001; 97(10): 3205-9 https://doi.org/10.1182/blood.V97.10.3205
  11. Wouters MGAJ, Boers GHJ, Blom HJ, Trijbels FJM, Tomas CMG, Borm GF. Hyperhomocysteinaemia: a risk factorin women with unexplained recurrent early pregnancy loss. Fertil Steril 1993; 60: 820-5
  12. Quere I, Bellet H, Hofet M, Janbon C, Mares P, Gris JC. A women with five consecutive fetal deaths: case report and retrospective analysis of hyperhomo- cysteinaemia prevalence in 100 consecutive women with recurrent miscarriages. Fertil Steril 1998; 68: 152-4
  13. Kawakami K, Omura K, Kanehira E, Watanabe Y. Polymorphic tandem repeats in the thymidylate syn- thase gene is associated with its protein expression in human gastrointestinal cancers. Anticancer Res 1999; 19: 3249-52
  14. Horie N, Aiba H, Oguro K, Hojo H, Takeishi K. Functionalanalysis and DNA polymorphism of the tandemly repeted sequences in the 5'-terminal regu- latory region of the human gene for thymidylate synthase. Cell Struct Funct 1995; 20: 191-197 https://doi.org/10.1247/csf.20.191
  15. Trinh BN, Ong C-N Goetzee GA, Yu MC, Laird PW. Thymidylate synthase: a novel genetic determinant of plasma homocysteine and folate levels. Hum Genet 2002; 111: 299-302 https://doi.org/10.1007/s00439-002-0779-2
  16. Kim NK, Nam YS, Lee S, Kim SH, Shin SJ, Chang SW, et al. Polymorphisms of 5,10-Methylenetetra- hydrofolate reductase (MTHFR C677T and A1298C) gene in recurrent spontaneous abortion. Kor J Fertil Steril 2002; 29(3): 215-222
  17. Kuferminc MJ, Eldor A, Steinman N, Many A, Bar-Am A, Jaffa A, et al. Increased frequency of genetic thrombophilia in women with complications of pregnancy. Lancet 1999; 340(1): 9-13 https://doi.org/10.1016/0140-6736(92)92423-D
  18. Kutteh WH, Park VM, Deither SR. Hypercoagu- lable state mutation analysis in white patients with early first-trimester recurrent pregnancy loss. Fertil Steril 1999; 71(6): 1048-53 https://doi.org/10.1016/S0015-0282(99)00133-8
  19. Brenner B, Sarig G, Weiner Z, Younis J, Blumenfeld Z, Lanir N. Thrombophilic polymorphisms are com- mon in women with fetal loss without apparent cause. Thromb Haemost 1999; 82(1): 6-9
  20. Nelen WLDM, Blom HJ, Steegers EAP, Thomas CMG, Boers GHJ, Eskes TKAB. Methylentetrahy- drofolate reductase polymorphism affects the change in homocysteine and folate concentration resulting from low dose folic acid supplementation in women unexplained recurrent miscarriages. J Nutr 1998; 128: 1336-41
  21. Nappo F, De Rosa N, Marfella R, De lucia D, In- grosso D, Perna AF, et al. Impairment of endothelial functions by acute hyperhomocysteinemia and re- versal by antioxidnt vitamins. JAMA 1999; 281(22): 2113-8 https://doi.org/10.1001/jama.281.22.2113
  22. Brown KS, Kluijtmans LA, Young IS, McNulty H, Mitchell LE, Yarnell JW, et al. The thymidylate synthase tandem repeat polymorphism is not asso- ciated with homocysteine concentrations in health young subjects. Hum Gent 2004; 114(2): 182-5 https://doi.org/10.1007/s00439-003-1039-9
  23. Ubbink J, Vermaak W, Van der Merue A, Becker P, Delport R, Potgeiter H. Vitaman requirements for the treatment of hyperhomocysteinaemia in humans. J Nutr 1994; 124: 1927-33
  24. Boushey CJ, Beresford SA, Omenn GS, Motulsky AG. A quqntytative assessment of plasma homocy- steine as a risk factor for vascular disease: probable benefits of increasing folic acid intakes. JAMA 1995; 274: 1049-57 https://doi.org/10.1001/jama.274.13.1049
  25. Den Heijer M, Rosendaal FR, Blom HJ, Gerrits WBJ, Bos GM. Hyperhomocystemia and venous thrombosis; a meta-analysis. Thromb Haemost 1998; 80: 874-7
  26. Obwegeser R, Hohlagschwandtner M, Sinzinger H. Homocysteine a pathophysiological cornerstone in obsterical and gynecological disorders? Hum Re- prod Update 1999; 5: 64-72
  27. French AE, Grant R, Weitzman S, Ray JG, Vermeu- len MJ, Sung L, et al. Folic acid food fortification is assocoated with a decline in neuroblastoma. Clin Pharmacol Ther 2003; 74(3): 288-94 https://doi.org/10.1016/S0009-9236(03)00200-5