臺灣博碩士論文加值系統

中文摘要
甘胺酸氮甲基轉化酶(glycine N-methyltransferase, 簡稱GNMT)，甲烯基四氫葉酸還原酶(5,10-methylenetetrahydrofolate reductase, 簡稱MTHFR)及甲硫胺酸生成酶 (Methionine synthase, 簡稱MS)主要參與了細胞單碳循環代謝路徑 (one carbon metabolism pathway)。DNA甲基化的程度會影響細胞染色體的穩定性及基因的表現，被認為與腫瘤的形成有關。由於以上個酵素皆會影響DNA甲基化的程度，所以本研究的目的有三：1. 探討GNMT，MTHFR和MS這3個基因的多型性與得到肝癌的相關性；2. 分析正常人血清GNMT濃度與其基因型的關係；3. 研究血清GNMT濃度的變化是否可以作為肝癌的一個臨床標記。
本研究收集183位肝癌病人與384位健康族群之週邊血液淋巴球（PBL），萃取其DNA，利用TaqMan及GeneScan方法鑑定GNMT的三個基因型（SNP1, STRP1及INS-DEL），以及利用PCR-RFLP方法定出MTHFR C677T及MS A2756G之基因型。結果發現肝癌病例組與健康對照組其GNMT，MTHFR、 MS的基因型分佈並無明顯差異。而在GNMT及MS的基因型的交互作用，發現在帶有 GNMT SNP1-C/C及 INS-DEL為DEL/DEL的人當中，若其PBL MS基因型含有G對偶基因（G/G 及 G/A型），則其罹患肝癌的風險為MS基因型為A/A 的2.33倍 (95% CI=1.19 - 4.59, p =0.014)；若其肝臟非腫瘤組織DNA檢體，GNMT INS-DEL為INS/DEL，其MS基因型含有G對偶基因，則其罹患肝癌的風險為MS基因型為A/A的5.08倍 (95% CI =1.27-20.32, p =0.015)。此外，肝癌病人的肝臟非腫瘤組織DNA檢體與健康對照組PBL比較，在MTHFR C/C型的人當中， MS 具有G對偶基因的較A/A基因型的人具有2.65倍罹患肝癌的風險 (95% CI = 1.02-6.86, p =0.041)。
進一步比較MTHFR、MS基因多型性在50對肝癌病人配對的腫瘤組織與非腫瘤組織DNA檢體中基因型的分佈，發現MTHFR基因體細胞DNA突變率為16.3%；MS基因體細胞DNA突變率則為10.0%。
最後，我們利用酵素免疫分析法針對452位正常人，195位慢性肝炎病人，28位肝硬化病人，及156位肝癌病人的血清測試他們的GNMT 濃度。結果發現以上四組的平均GNMT濃度分別為14.86 ± 10.49，17.78 ± 5.22，16.79 ± 5.08，6.12 ± 4.94 μg/ml。肝癌患者GNMT的表現量較健康對照組低，且具統計顯著差異(t-test，p值為<0.0001 )。在基因型與表現型的相關性方面，GNMT 的SNP1的CC型的GNMT濃度顯著高於TT型的濃度（p =0.038）。
綜合以上結果，推測在肝癌癌化過程中，甲基代謝路徑重要酵素MTHFR及MS具有高比例LOH現象，這可能是造成腫瘤的原因之一。此外，GNMT的血清濃度或許可以作為肝癌的一個臨床標記。

Abstract
Abnormal DNA methylation may affect chromosomal stability and gene expression. Many studies have shown an association between abnormal DNA methylation and an increased risk for the development of cancer. It has been shown that genes participated in the one carbon metabolism pathway, such as glycine N-methyltransferase (GNMT), 5, 10-methylenetetrahydrofolate reductase (MTHFR) and methionine synthase (MS) are involved in the regulation of the methylation level of DNA. Therefore, the goals of this study were 1. To tudy the association and interactions of the polymorphisms of genes mentioned above and the development of hepatocellular carcinoma (HCC); 2. To analyze the genotypic and phenotypic association of the plasma level of GNMT and the polymorphisms of mentioned above; 3. To evaluate whether the genotype of GNMT can be a diagnostic marker for HCC.
We analyzed the polymorphisms of the GNMT (SNP1, STRP1 and INS/DEL), MS (A2756G) and MTHFR (C677T) on the lymphocyte DNA obtained from 183 HCC patients and 384 normal controls. The results showed that the rates of each polymorphism among the above two groups were not significantly different. However, among people with GNMT SNP1-C/C + INS-DEL DEL/DEL, if their PBL MS genotype has G allele, they will have 2.33-fold risk (95% confidence interval [CI], 1.19-4.59, p =0.014) to develop HCC than subjects whose MS were A/A genotype; if their non-tumorous tissue with GNMT INS-DEL INS/DEL, MS genotype has G allele, then, they will have 5.08-fold risk than people’s non-tumorous tissue MS gene are A/A genotype (95% CI =1.27-20.32, p =0.015)。. In addition, among subjects with MTHFR C/C genotype, if their HCC non-tumorous tissues carrying MS G allele, they will have 2.65-fold risk for HCC than subjects whose non-tumorous tissues’ MS gene are A/A genotype (95% CI: 1.02-6.86, p =0.041).
In addition, among 50 pairs tumorous and non-tumorous tissues from HCC patients, the somatic mutation rates for MTHFR C677T and MS A2756G were 16.3%, 10.0%, respectively.
Blood samples from 452 normal adults, 195 patients with chronic hepatitis, 28 patients with cirrhosis and 156 patients with HCC were collected and their plasma level of GNMT were determined using an enzyme immunoassay. The results showed that the mean (± standard deviation) levels of GNMT were 14.86 ± 10.49，17.78 ± 5.22，16.79 ± 5.08 and 6.12 ± 4.94 μg/ml, respectively for those groups mentioned above and the differences were statistically significantly (ANOVA, p< 0.0001). In addition, among normal individuals, persons with GNMT SNP1 C/C genotype had significantly higher level of GNMT than persons with TT genotype; persons with MTHFR T allele had significantly higher levels of GNMT than persons with C/C genotype (p =0.038).
In conclusion, the interactions of the genetic polymorphisms of GNMT, MTHFR and MS, both in the PBL and non-tumorous liver tissues DNA, were associated with the risk of HCC. The plasma levels of GNMT correlate with certain genotypes of both GNMT and MTHFR genes and it has potential as a diagnostic marker for HCC.

參考文獻
Adami HO, Hsing AW, McLaughlin JK. Alcoholism and liver cirrhosis in the etiology of primary liver cancer. Int J Cancer 1992;51:898-902.
Amitrano L, Brancaccio V, Guardascione MA, Margaglione M, Iannaccone L, D'Andrea G, Marmo R, Ames PR, Balzano A. Inherited coagulation disorders in cirrhotic patients with portal vein thrombosis. Hepatology. 2000 Feb;31(2):345-8.
Antequera F, Bird A. Number of CpG islands and genes in human and mouse. Proc Natl Acad Sci USA 1993;90:11995-11999.
Austin H, Delzell E, Grufferman S, Levine R, Morrison AS, Stolley PD, Cole P. A case-control study of hepatocellular carcinoma and the hepatitis B virus, cigarette smoking, and alcohol consumption. Cancer Res 1986;46: 962-966.
Bagley PJ, Selhub J. A common mutation in the methylenetetrahydrofolate reductase gene is associated with an accumulation of formylated tetrahydrofolates in red blood cell. Proc Natl Acad Sci USA 1998;95:13217- 13220.
Balaghi M, Horne DW, Wagner C. Hepatic one-carbon metabolism in early folate deficiency in rats. Biochem J. 1993 Apr 1;291 ( Pt 1):145-9.
Baylin SB, Herman JG, Graff JR, Vertino PM, Issa JP. Alterations in DNA methylation: a fundamental aspect of neoplasia. Adv Cancer Res 1998;72: 141-196.
Beasley, R.P. Hepatitis B virus as the etiologic agent in hepatocellular carcinoma-
Epidemiologic considerations. Hepatology 1982; 2(suppl): 21-26.
Beasley RP. Hepatitis B virus: The major etiology of hepatocellular carcinoma. Cancer 1988; 61: 1942-1956.
Bhat R, Bresnick E. Glycine N-methyltransferase is an example of functional diversity. Role as a polycyclic aromatic hydrocarbon-binding receptor. J Biol Chem. 1997 Aug 22;272(34):21221-6.
Blount BC, Mack MM, Wehr CM, MacGregor JT, Hiatt RA, Wang G, Wickramasinghe SN, Everson RB, Ames BN. 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; 94: 3290-3295.
Bonassi S, Hagmar L, Stromberg U, Montagud AH, Tinnerberg H, Forni A, Heikkila P, Wanders S, Wilhardt P, Hansteen IL, Knudsen LE, Norppa H. Chromosomal aberrations in lymphocytes predict human cancer independently of exposure to carcinogens. European Study Group on Cytogenetic Biomarkers and Health. Cancer Res. 2000; 60: 1619-1625.
Chen CJ, Liang KY, Chang AS, Chang YC, Lu SN, Liaw YF, Chang WY, Sheen MC, Lin TM. Effects of hepatitis B virus, alcohol drinking, cigarette smoking and familial tendency on hepatocellular carcinoma. Hepatology 1991;13:398-406.
Chen CJ, Yu MW, Wang CJ, Huang HY, Lin WC. Multiple risk factors of hepatocellular carcinoma: A cohort study of 13,737 male adults in Taiwan. J Gastroenterol & Hepatol 1993;8:s83-s87.
Chen CJ, Yu MW, Liaw YF. Epidemiological characteristics and risk factors of hepatocellular carcinoma. J Gastroenterol & Hepatol 1997;12:S294-S308.
Chen DS, Kuo G, Sung JL, Lai MY, Sheu JC, Chen PJ, Yang PM, Hsu HM, Chang MH, Chen CJ, Hahn LC, Choo QL, Wang TH, Houghton M. Hepatitis C virus infection in an area hyperendemic for hepatitis B and chronic liver disease: The Taiwan experience. J Infect Dis 1990;162:817-822.
Chen J, Giovannucci E, Kelsey K, Rimm EB, Stampfer MJ, Colditz GA, Spiegelman D, Willett WC, Hunter DJ. A methylenetetrahydrofolate reductase polymorphism and the risk of colorectal cancer. Cancer Res, 1996; 56: 4862-4864.
Chen SY, Lin JR, Darbha R, Lin P, Liu TY, Chen YM. Glycine N-methyltransferase tumor susceptibility gene in the benzo(a)pyrene-detoxification pathway. Cancer Res. 2004 May 15;64(10):3617-23.
Chen YM, Shiu JY, Tzeng SJ, Shih LS, Chen YJ, Lui WY, Chen PH. Characterization of glycine-N-methyltransferase-gene expression in human hepatocellular carcinoma. Int J Cancer. 1998 Mar 2;75 (5):787-93.
Chen YM, Chen LY, Wong FH, Lee CM, Chang TJ, Yang-Feng TL. Genomic structure, expression, and chromosomal localization of the human glycine N-methyltransferase gene. Genomics. 2000 May 15;66(1):43-7.
Choi SW, Mason JB. Folate and carcinogenesis: an integrated scheme. J Nutr. 2000 Feb;130(2):129-32. Review.
Chung WL, Chang WY, Lu SN, Su WP, Lin ZY, Chen SC, Hsieh MY, Wang LY, You SL, Chen CJ. The role of hepatitis B and C viruses in hepatocellular carcinoma in a hepatitis B endemic area: A case-control study. Cancer 1992;69:2052-2054.
Cross SH, Bird AP. CpG islands and genes. Curr Biol 1995;5:309-314.
Duthie SJ. Folic acid deficiency and cancer: mechanisms of DNA instability. Br Med Bull. 1999;55(3):578-92. Review.
Fearon ER, Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990 Jun 1;61(5):759-67. Review.
Ferre N, Gomez F, Camps J, Simo JM, Murphy MM, Fernandez-Ballart J, Joven J. Plasma homocysteine concentrations in patients with liver cirrhosis. Clin Chem. 2002; 48: 183-185.
Frosst P, Blom HJ, Milos R, Goyette P, Sheppard CA, Matthews RG, Boers GJ, den Heijer M, Kluijtmans LA, van den Heuvel LP. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet 1995;10:111-113.
Fu Z, Hu Y, Konishi K, Takata Y, Ogawa H, Gomi T, Fujioka M, Takusagawa F. Crystal structure of glycine N-methyltransferase from rat liver. Biochemistry. 1996 Sep 17;35(37):11985-93.
Fuso A, Cavallaro RA, Orru L, Buttarelli FR, Scarpa S. Gene silencing by S-adenosylmethionine in muscle differentiation. FEBS Lett. 2001 Nov 23;508(3):337-40.
Gemmati D, Ongaro A, Scapoli GL, Della Porta M, Tognazzo S, Serino ML, Di Bona E, Rodeghiero F, Gilli G, Reverberi R, Caruso A, Pasello M, Pellati A, De Mattei M. Common gene polymorphisms in the metabolic folate and methylation pathway and the risk of acute lymphoblastic leukemia and non-Hodgkin's lymphoma in adults. Cancer Epidemiol Biomarkers Prev. 2004 May;13(5):787-94.
Giovannucci E, Stampfer MJ, Colditz GA, Hunter DJ, Fuchs C, Rosner BA, Speizer FE, Willett WC. Multivitamin use, folate, and colon cancer in women in the Nurses' Health Study. Ann Intern Med. 1998 Oct 1;129(7):517-24.
Glynn SA, Albanes D. Folate and cancer: a review of the literature. Nutr Cancer. 1994;22(2):101-19. Review.
Goyette P, Sumner JS, Milos R, Duncan AMV, Rosenblatt DS, Matthews RG, Rozen R. Human methylenetetrahydrofolate reductase: isolation of cDNA, mapping and mutation identification. Nat Genet 1994;7:195-200.
Hagmar L, Bonassi S, Stromberg U, Brogger A, Knudsen LE, Norppa H, Reuterwall C. Chromosomal aberrations in lymphocytes predict human cancer: a report from the European Study Group on Cytogenetic Biomarkers and Health (ESCH). Cancer Res. 1998; 58: 4117-4121.
Hann HL, Stahlhut MW, Blumberg BS. Iron nutrition and tumor growth: Decreased tumor growth in iron deficient mice. Cancer Res. 1988; 48: 4168-4170.
Hardell L, Bengtsson NO, Jonsson U, Eriksson S, Larsson LG. Aetiological aspects on primary liver cancer with special regard to alcohol, organic solvents and acute intermittent porphyria-an epidemiological investigation. Br J Cancer 1984;50:389-397.
Harmon DL, Shields DC, Woodside JV, McMaster D, Yarnell JW, Young IS, Peng K, Shane B, Evans AE and Whitehead AS. Methionine synthase D919G polymorphism is a significant but modest determinant of circulating homocysteine concentrations. Genetic Epidemiology. 17 (4): 298-309, 1999.
Hepatocellular cancer: differences between high and low incidence regions. Lancet. 1987 Nov 21;2(8569):1183-4. Review.
Hobbs CA, Sherman SL, Yi P, Hopkins SE, Torfs CP, Hine RJ, Pogribna M, Rozen R, James SJ. Polymorphisms in genes involved in folate metabolism as maternal risk factors for Down syndrome. Am J Hum Genet. 2000 Sep;67(3):623-30. Epub 2000 Aug 07.
Hsu HM, Chen DS, Chuang CH, Lu CF, Jwo DM, Lee CC, Lu HC, Cheng SH, Wang YF, Wang CC, Lo KJ, Shih CJ, Sung JL. Efficacy of a mass hepatitis B vaccination program in Taiwan: Studies on 3464 infants of hepatitis B surface-antigen-carrier mothers. JAMA 1988; 260: 2231-2235.
Idilman R, De Maria N, Colantoni A, Van Thiel DH. Pathogenesis of hepatitis B and C-induced hepatocellular carcinoma. J Viral Hepat 1998;5:285-299.
Issa JP, Ottaviano YL, Celano P, Hamilton SR, Davidson NE, Baylin SB. Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon. Nat Genet. 1994 Aug;7(4):536-40.
Jacob RA, Gretz DM, Taylor PC, James SJ, Pogribny IP, Miller BJ, Henning SM, Swendseid ME. Moderate folate depletion increases plasma homocysteine and decreases lymphocyte DNA methylation in postmenopausal women. J Nutr. 1998; 128: 1204-1212.
Kawakami K, Ruszkiewicz A, Bennett G, Moore J, Watanabe G, Iacopetta B. The folate pool in colorectal cancers is associated with DNA hypermethylation and with a polymorphism in methylenetetrahydrofolate reductase. Clin Cancer Res. 2003 Dec 1;9(16 Pt 1):5860-5.
Keen P, Martin P. Is aflatoxin carcinogenic in man? The evidence in Swaziland. Trop Geogr Med 1971;23:44-53.
Kim YI. Methylene-tetrahydrofolate reductase polymorphisms, folate, and cancer risk: a paradigm of gene-nutrient interactions in carcinogenesis. Nutr Rev 2000;58:205-209.
Kimura F, Florl AR, Steinhoff C, Golka K, Willers R, Seifert HH and Schulz WA. Polymorphic methyl group metabolism genes in patients with transitional cell carcinoma of the urinary bladder. [Journal Article] Mutation Research. 458 (1-2): 49-54, 2001.
Leclerc D, Campeau E, Goyette P, Adjalla CE, Christensen B, Ross M, Eydoux P, Rosenblatt DS, Rozen R and Gravel RA. Human methionine synthase: cDNA cloning and identification of mutations in patients of the cblG complementation group of folate/cobalamin disorders. Human Molecular Genetics. 5 (12): 1867-1874, 1996.
Lee, P.H., Lin, W.J., Tsang, Y.M., Hu, R.H., Sheu, J.C., Lai, M.Y., Hsu, H.C., May, W., and Lee, C.S. Clinical management of recurrent hepatocellular carcinoma. Ann. Surg 222: 670-676, 1995.
Lee SD, Lee FY, Wu JC, Hwang SJ, Wang SS, Lo KJ. The prevalence of anti-hepatitis C virus among Chinese patients with hepatocellular carcinoma. Cancer 1992;69:342-345.
Lengauer C, Kinzler KW, Vogelstein B. DNA methylation and genetic instability in colorectal cancer cells. Proc Natl Acad Sci USA 94: 2545-50, 1997.
Lin J, Spitz MR, Wang Y, Schabath MB, Gorlov IP, Hernandez LM, Pillow PC, Grossman HB, Wu X. Polymorphisms of folate metabolic genes and susceptibility to bladder cancer: a case-control study. Carcinogenesis. 2004 Apr 29
Lu SN, Lin TM, Chen CJ, Chen TS, Liaw YF, Chang WY, Hsu ST. A case-control study of primary hepatocellular carcinoma in Taiwan. Cancer 1988;62:2051-2055.
Luka Z, Cerone R, Phillips JA 3rd, Mudd HS, Wagner C. Mutations in human glycine N-methyltransferase give insights into its role in methionine metabolism. Hum Genet. 2002 Jan;110(1):68-74. Epub 2001 Dec 07.
Ma J, Stampfer MJ, Hennekens CH, Frosst P, Selhub J, Horsford J, Malinow MR, Willett WC, Rozen R. Methylenetetrahydrofolate reductase polymorphism, plasma folate, homocysteine, and risk of myocardial infarction in US physicians. Circulation 1996; 94:2410-2416.
Ma J, Stampfer MJ, Giovannucci E, Artigas C, Hunter DJ, Fuchs C, Willett WC, Selhub J, Hennekens CH, Rozen R. Methylenetetrahydrofolate reductase polymorphism, dietary interactions, and risk of colorectal cancer. Cancer Res, 1997; 57: 1098-1102.
Moore LE, Wiencke JK, Bates MN, Zheng S, Rey OA, Smith AH. Investigation of genetic polymorphisms and smoking in a bladder cancer case-control study in Argentina. Cancer Lett. 2004 Aug 10;211(2):199-207.
Morita H, Taguchi J, Kurihara H, Kitaoka M, Kaneda H, Kurihara Y, Maemura K, Shindo T, Minamino T, Ohno M, Yamaoki K, Ogasawara K, Aizawa T, Suzuki S, Yazaki Y. [Gene Polymorphism of 5, 10-methylenetetrahydrofolate reductase as a coronary risk factor] J Cardiol. 1997 Jun;29(6):309-15. Japanese.
Morita H, Kurihara H, Sugiyama T, Hamada C, Kurihara Y, Shindo T, Oh-hashi Y and Yazaki Y. Polymorphism of the methionine synthase gene: Association with homocysteine metabolism and late-onset vascular diseases in the Japanese population. Arteriosclerosis, Thrombosis & Vascular Biology. 19 (2): 298-302, 1999.
Mu LN, Ding BG, Chen CW, Wei GR, Zhou XF, Wang RH, Cai L, Zhang ZF, Jiang QW, Yu SZ. [A case-control study on the relationship between methyl-tetra-hydrofolic acid reductase 677 gene polymorphism and the risk of stomach cancer] Zhonghua Liu Xing Bing Xue Za Zhi. 2004 Jun;25(6):495-8. Chinese.
Mudd SH, Cerone R, Schiaffino MC, Fantasia AR, Minniti G, Caruso U, Lorini R, Watkins D, Matiaszuk N, Rosenblatt DS, Schwahn B, Rozen R, LeGros L, Kotb M, Capdevila A, Luka Z, Finkelstein JD, Tangerman A, Stabler SP, Allen RH, Wagner C. Glycine N-methyltransferase deficiency: a novel inborn error causing persistent isolated hypermethioninaemia. J Inherit Metab Dis. 2001 Aug;24(4):448-64.
Nishimura M, Yoshino K, Tomita Y, Takashima S, Tanaka J, Narisawa K, Kurobane I. Central and peripheral nervous system pathology of homocystinuria due to 5,10-methylenetetrahydrofolate reductase deficiency. Pediatr Neurol. 1985 Nov-Dec;1(6):375-8.
Ogawa H, Gomi T, Fujioka M. Mammalian glycine N-methyltransferases. Comparative kinetic and structural properties of the enzymes from human, rat, rabbit and pig livers. Comp Biochem Physiol B. 1993 Nov;106(3):601-11.
Parkin DM, Pisani P, Ferlay J. Estimates of the worldwide incidence of eighteen major cancers in 1985. Int J Cancer. 1993 Jun 19;54(4):594-606
Poirier LA. Methyl group deficiency in hepatocarcinogenesis. Drug Metab Rev. 1994;26(1-2):185-99. Review.
Patrizia DM, Maria GC, Anna M, Lorenza A, Elisa M, Richard HF, Huiping Z, Luciano A, Armando C, Valeria C. Study of MTHFR and MS polymorphisms as risk factors for NTD in the Italian population. J Hum Genet (2002) 47:319-324.
Poland A, Glover E, Kende AS. J. Biol. Chem 1976; 251-4936-4946.
Qian GS, Ross RK, Yu MC, Yuan JM, Gao YT, Henderson BE, Wogan GN, Groopman JD. A follow-up study of urinary markers of aflatoxin exposure and liver cancer risk in Shanghai, People’s Pepublic of China. Cancer Epidemiol Biomark & Prev 1994;3:3-10.
Qu GZ, Grundy P, Narayan A, Ehrlich M. Frequent hypomethylation in Wilms tumors of pericentromeric DNA in chromosomes 1 and 16. Cancer Genet Cytogenet 76: 196-201, 1998.
Raha A, Wagner C, MacDonald RG, Bresnick E. Rat liver cytosolic 4 S polycyclic aromatic hydrocarbon-binding protein is glycine N-methyltransferase. J Biol Chem. 1994 Feb 25;269(8):5750-6.
Rhee I, Bachman KE, Park BH, Jair KW, Yen RW, Schuebel KE, Cui H, Feinberg AP, Lengauer C, Kinzler KW, Baylin SB, Vogelstein B. DNMT1 and DNMT3b cooperate to silence genes in human cancer cells. Nature. 2002 Apr 4;416(6880):552-6.
Rowling MJ, Schalinske KL. Retinoid compounds activate and induce hepatic glycine N-methyltransferase in rats. J Nutr. 2001 Jul;131(7):1914-7.
Rowling MJ, McMullen MH, Schalinske KL. Vitamin A and its derivatives induce hepatic GNMT and hypomethylation of DNA in rats. J. Nutr. 2002; 132:365-369.
Saffroy R, Pham P, Chiappini F, Goupil MG, Castera L, Azoulay D, Barrier A, Samuel D, Debuire B, Lemoine A. The MTHFR 677C>T polymorphism is associated with an increased risk of hepatocellular carcinoma in patients with alcoholic cirrhosis. Carcinogenesis. 2004 Mar 19
Schwahn B and Rozen R. Polymorphisms in the methylenetetrahydrofolate reductase gene. Am J Pharmacogenomics 2001;1:189-201.
Sibani S, Christensen B, O’Ferrall E, Saadi I, Hiou-Tim F, Rosenblatt DS, Rozen R. Characterization of six novel mutations in the methylenetetrahydrofolate reductase (MTHFR) gene in patients with homocystinuria. Hum Mutat 2000;15:280-287.
Simonetti RG, Camma C, Fiorello F, Politi F, D'Amico G, Pagliaro L. Hepatocellular carcinoma. A worldwide problem and the major risk factors. Dig Dis Sci. 1991 Jul;36(7):962-72. Review.
Singal R, Ginder GD. DNA methylation. Blood 1999;93:4059-4070.
Slattery ML, Potter JD, Samowitz W, Schaffer D, Leppert M Methylenetetrahydrofolate reductase, diet, and risk of colon cancer. Cancer Epidemiol. Biomarkers Prev, 1999; 8: 513-518.
Stern, L. L., Mason, J. B., Selhub, J., and Choi, S. W. Genomic DNA hypomethylation, a characteristic of most cancers, is present in peripheral leukocytes of individuals who are homozygous for the C677T polymorphism in the methylenetetrahydrofolate reductase gene. Cancer Epidemiol. Biomark. Prev., 9: 849–853, 2000.
Swanson DA, Liu ML, Baker PJ, Garrett L, Stitzel M, Wu J, Harris M, Banerjee R, Shane B and Brody LC. Targeted disruption of the methionine synthase gene in mice. [Journal Article] Molecular & Cellular Biology. 21 (4): 1058-1065, 2001.
Tanaka K, Hirohata T, Takeshita S, Hirohata I, Koga S, Sugimachi K, Kanematsu T, Ohryohji F, Ishibashi H. Hepatitis B virus, cigarette smoking and alcohol consumption in the development of hepatocellular carcinoma: a case-control study in Fukuoka, Japan. Int J Cancer 1992;51:509-514.
Tierney B, Weaver D, Heintz NH, Schaeffer WI, Bresnick E. The identity and nuclear uptake of a cytosolic binding protein for 3-methylcholanthrene. Arch Biochem Biophys. 1980 Apr 1;200(2):513-23.
Trimble K. C., Molloy A. M., Scott J. M., Weir D. G. The effect of ethanol on one-carbon metabolism: increased methionine catabolism and lipotrope methyl-group wastage. Hepatology 1991;18:984-989
Tsai JF, Chuang LY, Jeng JE, Ho MS, Hsieh MY, Lin ZY, Wang LY. Betel quid chewing as a risk factor for hepatocellular carcinoma: a case-control study. Br J Cancer 2001;84:709-713.
Tsai MY, Bignell M, Schwichtenberg K and Hanson NQ. High prevalence of a mutation in the cystathionine beta-synthase gene. American Journal of Human Genetics. 59 (6): 1262-1267, 1996.
Tseng TL, Shih YP, Huang YC, Wang CK, Chen PH, Chang JG, Yeh KT, Chen YM, Buetow KH. Genotypic and phenotypic characterization of a putative tumor susceptibility gene, GNMT, in liver cancer. Cancer Res 63: 647-654, 2003
Tsukuma H, Hiyama T, Oshima A, Sobue T, Fujimoto I, Kasugai H, Kojima J, Sasaki Y, Imaoka S, Horiuchi N. A case-control study of hepatocellular carcinoma in Osaka, Japan. Int J Cancer 1990;45:231-236.
Van der Put NM, Steegers-Theunissen RP, Frosst P, Trijbels FJ, Eskes TK, van den Heuvel LP, Mariman EC, den Heyer M, Rozen R, Blom HJ. Mutated methylenetetrahydrofolate reductase as a risk factor for spina bifida. Lancet. 1995 Oct 21;346(8982):1070-1.
Van der Put NMJ, Gabreels F, Stevens EMB, Smeitink JAM, Trijbels FJM, Eskes TKAB, Van den Heuvel LP, Blom HJ. A second common mutation in the methylenetetrahydrofolate reductase gene: an additional risk factor for neural-tube defects? Am J Hum Genet 1998;62:1044-1051.
Ven den Veyver IB. Genetic effects of methylation diets. Ann Rev Nutr 2002;22: 255-82.
Walk D, Kang SS, Horwitz A. Intermittent encephalopathy, reversible nerve conduction slowing, and MRI evidence of cerebral white matter disease in methylenetetrahydrofolate reductase deficiency. Neurology. 1994 Feb;44(2):344-7.
Wagner C, Briggs WT, Cook RJ. Inhibition of glycine N-methyltransferase activity by folate derivatives: implicatioins for regulation of methyl group metabolism. Biochem Biophys Res Commun. 1985 Mar 29; 127 (3): 746-52.
Wagner C., Decha-Umphai W., Corbin J. Phosphorylation modulates the activity of glycine-N-methyltransferase, a folate binding protein: in vitro phosphorylation is inhibited by the natural folate ligand. J. Biol. Chem. 1989;264:9638-9642
Whitehead AS, Gallagher P, Mills JL, Kirke PN, Burke H, Molloy AM, Weir DG, Shields DC, Scott JM. A genetic defect in 5,10-methylenetetrahydrofolate reductase in neural tube defects. Q. J. Med. 88: 763-766, 1995.
Yates Z, Lucock M. Methionine synthase polymorphism A2756G is associated with susceptibility for thromboembolic events and altered B vitamin/thiol metabolism.
Haematologica. 2002 Jul;87(7):751-6; discussion 756.
Yeo EJ, Wagner C. Tissue distribution of glycine N-methyltransferase, a major folate-binding protein of liver. Proc Natl Acad Sci U S A. 1994 Jan 4;91 (1):210-4.
Yu MC, Mack T, Hanisch R, Peters RL, Henderson BE, Pike MC. Hepatitis, alcohol consumption, cigarette smoking, and hepatocellular carcinoma in Los Angeles. Cancer Res 1983;43:6077-6079.
Yu MC, Tong MJ, Govindarajan S, Henderson BE. Nonviral risk factors for hepatocellular carcinoma in a low-risk population, the non-Asians of Los Angeles County, California. J Natl Cancer Inst 1991;83:1820-1826.
Yu MW, Tsai SF, Hsu KH, You SL, Lee SS, Lin TM, Chen CJ. Epidemiologic characteristics of malignant neoplasms in Taiwan:Ⅱ. Liver cancer. J Natl Public Health Assoc (ROC) 1988;8:125-138.
Yu MW, You SL, Chang AS, Lu SN, Liqw YF, Chen CJ. Association between hepatitis C virus antibodies and hepatocelular carcinoma in Taiwan. Cancer Res. 51:5621-5625, 1991.
Yu MW, Chen CJ. Elevated serum testosterone levels and risk of hepatocellular carcinoma. Cancer Res 1993;53:790-794.
Yu MW, Hsieh HH, Pan WH, Yang CS, Chen CJ. Vegetable consumption, serum retinal level, and risk of hepatocellular carcinoma. Cancer Res 1995a;55:1301-1305.
Yu MW, Chang HC, Liaw YF, Lin SM, Lee SD, Liu CJ, Chen PJ, Hsiao TJ, Lee PH, Chen CJ. Familial risk of hepatocellular carcinoma among chronic hepatitis B carriers and their relatives. J Natl Cancer Inst 2000a;92:1159-1164.
Zhang GS and Dai CW Gene polymorphisms of homocysteine metabolism-related enzymes in Chinese patients with occlusive coronary artery or cerebral vascular diseases. Thrombosis Research. 104 (3): 187-195, 2001.
Zhu WL, Cheng J, Dao JJ, Zhao RB, Yan LY, Li SQ, Li Y. Polymorphism of methionine synthase gene in nuclear families of congenital heart disease. Biomed Environ Sci. 2004 Mar;17(1):57-64.
Zingg JM, Jones PA. Genetic and epigenetic aspects of DNA methylation on genome expression, evolution, mutation and carcinogenesis. Carcinogenesis 1997;18:869-882.
行政院衛生署。中華民國八十九年癌症登記報告。行政院衛生署，2000 年。
行政院衛生署。中華民國九十二年生命統計資料。行政院衛生署，2003 年。
行政院衛生署。中華民國九十二年國內死亡原因表。行政院衛生署，2003 年。