摘要
研究目的:
在江苏省宜兴市进行以人群为基础的分子流行病学病例-对照研究,筛选错配修复基因hMLH1和hMSH2有功能意义的4个SNPs,探讨hMLH1和hMSH2基因突变与我国胃癌发生风险的关联性。最终为阐明胃癌发病的分子遗传学机制,指导高危人群有针对性的预防和治疗提供依据。
研究方法:
以江苏省宜兴市554例胃癌患者和592例年龄(±5岁)、性别相匹配的非胃癌患者(对照组)作为研究对象,用TaqMan MGB探针对hMLH1基因-93G＞A、1219V和hMSH2基因-118T＞C、IVS12-6T＞C四个SNPs进行基因分型,χ2检验比较研究人群年龄、性别、吸烟、饮酒等因素频数分布差异,揭示胃癌发生与吸烟、饮酒等生活习惯之间的关系。非条件Logistic回归分析不同基因型与胃癌发生风险的关联性。分层分析探讨不同基因型与胃癌发生的性别、年龄、吸烟、饮酒及其临床病理特征之间的关联性。
结果:
1,胃癌病例中吸烟、饮酒者多于对照组,和对照组相比有显著性差异(P＜0.001和P=0.002)。
2,在病例-对照研究中我们发现hMLH1基因-93G＞A、1219V和hMSH2基因-118T＞C、IVS12-6T＞C四个SNPs与胃癌的发生风险无显著关联性(P＞0.05)。分层分析我们发现hMSH2基因-118T＞C合并突变基因型(TC+CC)和对照相比与年轻人群(年龄≤63岁)胃癌和弥漫型胃癌的发生风险有显著关联性(调整OR=1.51,95%CI=1.05-2.16和调整OR=1.41,95%CI=1.01-1.96)。而其余三个SNPs尚未发现此种关联性。
结论:
1,胃癌的发生是复杂的,多因素的,有许多已知的因素参与了胃癌的发生,如吸烟、饮酒等因素。
2,错配修复基因hMLH1-93G＞A、I219V和hMSH2IVS12-6T＞C多态可能与胃癌的发生风险无显著关联性。
3,hMSH2-118T＞C多态可增加年轻人群胃癌和弥漫型胃癌的发生风险。
Objective:
Population-based case-control study on molecular epidemiology was carried out in Yixing city, which in the Jiangsu Province of China. To assess the association between the mismatch repair genes hMLH1 and hMSH2 polymorphisms and the risk of gastric cancer in China, four polymorphic sites with functional significance of hMLH1 and hMSH2 genes were selected. In the end the mechanism of the molecular genetics which lead to the incidence of gastric cancer will be clarified. Furthermore, this study will guide the prevention and therapy for high-risk groups.
Methods:
554 patients with gastric cancer cases and 592 cancer-free controls frequency-matched by age (±5) and sex were recruited in the study. The genotypes of the hMLH1-93G>A, I219V and hMSH2-118T>C, IVS12-6T>C polymorphisms were detected by TaqMan MGB probe method. Chi-square test was used to evaluate differences in frequency distributions of age, sex, smoking status and alcohol use between the cases and controls and further to assess the association with the occurrence of gastric cancer. Unconditional logistic regression was used to assess the association between each polymorphism and gastric cancer. We further analyzed the interaction of age, sex, smoking, alcohol use and tumor clinic pathological characteristics.
Results:
1. Gastric cancer cases have more smokers and drinkers than controls and the association between them was statistically significant (P < 0.001 and P = 0.002, respectively).
2. No evidence of an association between any of the four polymorphisms and gastric cancer was observed (P > 0.05) . Further analysis revealed that, when compared with the control populations, the hMSH2-118 combined variant genotypes (TC+CC) were increased the risk of gastric cancer accompanied with younger subjects (age≤63 years; adjusted odds ratios [OR] = 1.51, 95% confidence intervals [CI] = 1.05-2.16) and associated with diffuse-type tumors (adjusted OR =1.41, 95% CI = 1.01-1.96).
Conclusion:
1. Gastric cancer is a complex and multifactorial process. There are many known factors involved in the occurrence of gastric cancer, such as smoking and alcohol use.
2. The hMLH1-93G>A, I219V and hMSH2 IVS12-6T>C polymorphisms may not contribute to risk of gastric cancer.
3. The hMSH2-118T>C polymorphism may confer not only an increased risk of younger subjects (age≤63 years) of gastric cancer but also with diffuse-type tumors.
引文
Parkin DM,Bray F,Ferlay J,et al.Global cancer statistics,2002[J].CA Cancer J Clin,2005,55(2):74-108.
2.周标,陈坤.胃癌主要危险因素研究进展[J].金华职业技术学院学报,2005,5(3):37-39.
3.Stadtlander CT,Waterbor JW.Molecular epidemiology,pathogenesis and prevention of gastric cancer[J].Carcinogenesis,1999,20(12):2195-2208.
4.Beales IL.H.pylori-associated hypochlorhydria[J].Gastroenterology,1998,114(3):618-621.
5.Lan J,Xiong YY,Lin YX,et al.Helicobacter pylori infection generated gastric cancer through p53-Rb tumor-suppressor system mutation and telomerase reactivation[J].World J Gastroenterol,2003,9(1):54-58.
6.Koessler T,Oestergaard MZ,Song H,et al.Common variants in mismatch repair genes and risk of colorectal cancer[J].Gut,2008,57(8):1097-1101.
7.Chen Y,Fraig MM,Wang J,et al.Defects of DNA mismatch repair in human prostate cancer[J].Cancer Res,2001,61(10):4112-4121.
8.Ilyas M,Straub J,Tomlinson IP,et al.Genetic pathways in colorectal and other cancers [J].Eur J Cancer,1999,35(14):1986-2002.
9.Lynch HT,de la Chapelle A.Hereditary colorectal cancer[J].N Engl J Med,2003,348(10):919-932.
10.Hampel H,Frankel WL,Martin E,Arnold M,Khanduja K,Kuebler P,et al.Screening for the Lynch syndrome(hereditary nonpolyposis colorectal cancer)[J].N Engl J Med,2005,352(18):1851-1860.
11.Chung DC,Rustgi AK.The hereditary nonpolyposis colorectai cancer syndrome:genetics and clinical implications[J].Ann Intern Med,2003,138(7):560-570.
12.Rowley PT.Inherited susceptibility to colorectal cancer[J].Annu Rev Med,2005,56:539-554.
13.Lynch HT,de la Chapelle A.Hereditary colorectal cancer[J].N Engl J Med,2003, 348(10):919-932.
14.Mitchell RJ,Farrington SM,Dunlop MG,et al.Mismatch repair genes hMLH1 and hMSH2 and colorectal cancer:a HuGE review[J].Am J Epidemiol,2002,156(10):885-902.
15.Lee KM,Choi JY,Kang C,et al.Genetic polymorphisms of selected DNA repair genes,estrogen and progesterone receptor status,and breast cancer risk[J].Clin Cancer Res,2005,11(12):4620-4626.
16.Beiner ME,Rosen B,Fyles A,et al.Endometrial cancer risk is associated with variants of the mismatch repair genes MLH1 and MSH2[J].Cancer Epidemiol Biomarkers Prey,2006,15(9):1636-1640.
17.Raptis S,Mrkonjic M,Green RC,et al.MLH1-93G＞A promoter polymorphism and the risk of microsatellite-unstable colorectal cancer[J].J Natl Cancer Inst,2007,99(6):463-474.
18.Smith TR,Levine EA,Freimanis RI,et al.Polygenic model of DNA repair genetic polyrnorphisms in human breast cancer risk[J].Carcinogenesis,2008,29(11):2132-2138.
19.Hishida A,Matsuo K,Hamajima N,et al.Polymorphism in the hMSH2 gene gIVS126T＞C and risk of non-Hodgkin lymphoma in a Japanese population[J].Cancer Genet Cytogenet,2003,147(1):71-74.
20.Jung CY,Choi JE,Park JM,et al.Polymorphisms in the hMSH2 gene and the risk of primary lung cancer[J].Cancer Epidemiol Biomarkers Prev,2006,15(4):762-768.
21.Mrkonjic M,Raptis S,Green RC,et al.MSH2-118T＞C and MSH6-159C＞T promoter polymorphisms and the risk of colorectal cancer[J].Carcinogenesis,2007,28(12):2575-2580.
22.王花丽.错配修复毖冈hMSH2、hMLH1基因表达与胃癌发生关系的研究[C].大连医科大学硕士学位论文,2006.
23.杨家红,范宗华等.吸烟、饮酒与肖癌死亡关系的前瞻性队列研究[J].现代预防医学,2000,27(3):322-324.
24.Kato I,Vivas J,Plummer M,et al.Environmental factors in Helicobacter pylori-related gastric precancerous lesions in Venezuela[J].Cancer Epidemiol Biomarkers Prey,2004,13(3):468-476.
25.Peleteiro B,Lunet N,Figueiredo C,et al.Smoking,Helicobacter pylori virulence,and type of intestinal metaplasia in Portuguese males[J].Cancer Epidemiol Biomarkers Prey,2007,16(2):322-326.
26.路长东,张栓虎.国内饮酒与胃癌关系的Meta分析[J].新乡医院院学报.2007,1(24):66-68.
27.Lynch HT,Lynch J.Lynch syndrome:genetics,natural history,genetic counseling and prevention[J].J Clin Oncol,2000,18(21Suppl):19S-31S.
28.Dunlip MG,Farrington SM,Carothers AD,et al.Cancer risk associated with germline DNA mismatch repair gene mutation[J].Hum Mol Genet,1997,6(1):105-110.
29.Ito E,Yanagisawa Y,Iwahashi Y,et al.A core promoter and a frequent single- nucleotide polymorphism of the mismatch repair gene hMLH1[J].Biochem Biophys Res Commun,1999,256(3):488-494.
30.Zenke M,Grundstrom T,Matthes H,et al.Multiple sequence motifs are involved in SV40enhancer function[J].EMBO.J,1986,5(2):387-97.
31.Kinoshita S,Akira S,Kishimoto T.A member of the C/EBP family,NF-IL6 beta,forms a heterodimer and transcriptionally synergizes with NF-IL6[J].Proc Natl Acad Sci U S A,1992,89(4):1473-1476.
32.Park SH,Lee GY,Jeon HS,et al.-93G＞A polymorphism of hMLH1 and risk of primary lung cancer[J].Int J Cancer,2004,112(4):678-682.
33.Song H,Ramus SJ,Quaye L,et al.Common variants in mismatch repair genes and risk of invasive ovarian cancer[J].Carcinogenesis,2006,27(11):2235-2242.
34.Campbell PT,Curtin K,Ulrich CM,et al.Mismatch repair polymorphisms and risk of colon cancer,tumour microsatellite instability and interactions with lifestyle factors[J].Gut,2009,58(5):661-667.
35.Bagnoli S,Putignano AL,Melean G,et al.Susceptibility to refractory ulcerative colitis is associated with polymorphism in the hMLH1 mismatch repair gene[J].Inflamm Bowel Dis,2004,10(6):705-708.
36.Mathonnet G,Krajinovic M,Labuda D,et al.Role of DNA mismatch repair genetic polymorphisms in the risk of childhood acute lymphoblastic leukaemia[J].Br J Haematol,2003,123(1):45-48.
37. Fishel R, Lescoe MK, Rao MR, et al. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer [J]. Cell, 1993, 75(5): 1027-1038.
38. Palicio M, Blanco I, Tortola S, et al. Intron splice acceptor site polymorphism in the hMSH2 gene in sporadic and familial colorectal cancer [J]. Br J Cancer, 2000, 82(3):535-537.
39. Shin KH, Shin JH, Kim JH, et al. Mutational analysis of promoters of mismatch repair genes hMSH2 and hMLH1 in hereditary nonpolyposis colorectal cancer and early onset colorectal cancer patients: identification of three novel germ-line mutations in promoter of the hMSH2 gene [J]. Cancer Res, 2002, 62(1):38-42.
40. Iwahashi Y, Ito E, Yanagisawa Y, et al. Promoter analysis of the human mismatch repair gene hMSH2 [J]. Gene, 1998, 213(1-2): 141-147.
41. Miyamoto T, Shiozawa T, Kashima H, et al. Estrogen up-regulates mismatch repair activity in normal and malignant endometrial glandular cells [J]. Endocrinology, 2006, 147(10):4863-4870.
42. Rosenberg NA, Pritchard JK, Weber JL, et al. Genetic structure of human populations [J]. Science, 2002, 298(5602):2381-2385.
43. Gu H, Yang L, Sun Q, et al. Gly82Ser polymorphism of the receptor for advanced glycation end products is associated with an increased risk of gastric cancer in a Chinese population [J]. Clin Cancer Res, 2008, 14(11):3627-3632.
44. Yang L, Gu HJ, Zhu HJ, et al. Tissue inhibitor of metalloproteinase-2 G-418C polymorphism is associated with an increased risk of gastric cancer in a Chinese population [J]. Eur J Surg Oncol, 2008, 34(6):636-641.
45. Zhu H, Yang L, Zhou B, et al. Myeloperoxidase G-463A polymorphism and the risk of gastric cancer: a case-control study [J]. Carcinogenesis, 2006, 27(12):2491-2496.
46. Sakamoto H, Yoshimura K, Saeki N, et al. Genetic variation in PSCA is associated with susceptibility to diffuse-type gastric cancer [J]. Nat Genet, 2008, 40(6):730-740.
1. Yashiro M, Carethers JM, Laghi L, et al. Genetic pathways in the evolution of morphologically distinct colorectal neoplasms [J]. Cancer Res, 2001, 61(6):2676-2683.
2. Jacob S, Praz F. DNA mismatch repair defects: role in colorectal carcinogenesis [J]. Biochimie, 2002, 84(1):27-47.
3. Raptis S, Mrkonjic M, Green RC, et al. MLH1 -93G>A promoter polymorphism and the risk of microsatellite-unstable colorectal cancer [J]. J Natl Cancer Inst, 2007, 99(6):463-474.
4. Lee KM, Choi JY, Kang C, et al. Genetic polymorphisms of selected DNA repair genes, estrogen and progesterone receptor status, and breast cancer risk [J]. Clin Cancer Res, 2005, 11(12):4620-4626.
5. Beiner ME, Rosen B, Fyles A, et al. Endometrial cancer risk is associated with variants of the mismatch repair genes MLH1 and MSH2 [J]. Cancer Epidemiol Biomarkers Prev, 2006, 15(9): 1636-1640.
6. Burdett V, Baitinger C, Viswanathan M, et al. In vivo requirement for RecJ, ExoVII, Exol, and ExoX in methyl-directed mismatch repair [J]. Proc Natl Acad Sci U S A, 2001, 98(12):6765-6770.
7. Hall MC, Matson SW. The Eschenchia coli MutL protein physically interacts with MutH and stimulates the MutH-associated endonuclease activity [J]. J Biol Chem, 1999, 274(3):1306-1312.
8. Lahue RS, Au KG, Modrich P. DNA mismatch correction in a defined system [J]. Science, 1989, 245(4914):160-164.
9. Lu AL, Clark S, Modrich P. Methyl-directed repair of DNA base-pair mismatches in vitro [J]. Proc Natl Acad Sci U S A, 1983, 80(15):4639-4643.
10. Cooper DL, Lahue RS, Modrich P. Methyl-directed mismatch repair is bidirectional [J]. J Biol Chem, 1993, 268(16): 11823-11829.
11. Fishel R, Lescoe MK, Rao MR, et al. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer [J]. Cell, 1993, 75(5): 1027-1038.
12. Dunlip MG, Farrington SM, Carothers AD, et al. Cancer risk associated with germline DNA mismatch repair gene mutation [J]. Hum Mol Genet, 1997, 6(1):105-110.
13. Kolodner RD, Richard, Nigee R, et al. Structure of the human MLH1 locus and analysis of large hereditary nonpolyposis colorectal carcinoma kinred for MHL1 mutation [J].Cancer Res, 1995,55:242
14. Peltomaki P. DNA mismatch repair and cancer [J]. Mutat Res, 2001, 488(1):77-85.
15. McCulloch SD, Gu L, Li GM. Bi-directional processing of DNA loops by mismatch repair-dependent and -independent pathways in human cells [J]. J Biol Chem, 2003, 278(6):3891-3896.
16. Aaltonen LA, Peltomaki P, Leach FS, et al. Clues to the pathogenesis of familial colorectal cancer [J]. Science, 1993, 260(5109):812-816.
17. Sinicrope FA, Rego RL,et al, Alterations in cell proliferation and apoptosis in colon cancers with microsatellite instability [J]. Int J Cancer. 2007 , 120(6): 1232-1238.
18. Heinen CD, Schmutte C, Fishel R. DNA repair tumor genesis lessons from hetediary cancer syndromes [J], Cancer Biol Ther, 2002, 1(5):477-485.
19. Eshleman JR, Lang EZ, Bowerfind GK, et al. Increased mutation rate at the hprt locus accompanies microsatellite instability in colon cancer [J]. Oncogene, 1995, 10(1):33-37.
20. Yan T, Berry SE, Desai AB, Kinsella TJ. DNA mismatch repair (MMR) mediates 6-thioguanine genotoxicity by introducing single-strand breaks to signal a G2-M arrest in MMR-proficient RKO cells [J]. Clin Cancer Res, 2003, 9(6):2327-2334.
21. Yokozaki H, Yasui W, Tahara E. Genetic and epigenetic changes in stomach cancer [J]. Int Rev Cytol, 2001, 204:49-95.
22. Koessler T, Oestergaard MZ, Song H, et al. Common variants in mismatch repair genes and risk of colorectal cancer [J]. Gut, 2008, 57(8):1097-1101.
23. Chung DC, Rustgi AK. The hereditary nonpolyposis colorectal cancer syndrome: genetics and clinical implications [J]. Ann Intern Med, 2003, 138(7):560-570.
24. Yashiro M, Carethers JM, Laghi L, et al. Genetic pathways in the evolution of morphologically distinct colorectal neoplasms [J]. Cancer Res, 2001, 61(6):2676-2683.
25. Mitchell RJ, Farrington SM, Dunlop MG, et al. Mismatch repair genes hMLH1 and hMSH2 and colorectal cancer: a HuGE review [J]. Am J Epidemiol, 2002, 156(10):885-902.
26.Gylling A,Abdel-Rahman WM,Juhola M,et al.Is gastric cancer part of the tumour spectrum of hereditary non-polyposis colorectal cancer? A molecular genetic study[J].Gut,2007,56(7):926-933.
27.Kim JC,Kim HC,Roh SA,et al.hMLH1 and hMSH2 mutations in families with familial clustering of gastric cancer and hereditary non-polyposis colorectal cancer[J].Cancer Detect Prev,2001,25(6):503-510.
28.王亚平,周建农,李忠佑.中国人消化道肿瘤发病的遗传背景因素[J].中华医学遗传学杂志,2000,17(2):82-86
29.李建华,石先哲,吕申,等.胃癌家系人群基因点突变的分析[J].大连医科大学学报.2004,26(4):241-243.
30.赵成海,刘宏旭,卜献民.错配修复基因hMLH3在家族性胃癌中的突变[J].世界华人消化杂志,2004,12(5):1030-1033.
31.Smith TR,Levine EA,Freimanis RI,et al.Polygenic model of DNA repair genetic polymorphisms in human breast cancer risk[J].Carcinogenesis,2008,29(11):2132-2138.
32.Jung CY,Choi JE,Park JM,et al.Polymorphisms in the hMSH2 gene and the risk of primary lung cancer[J].Cancer Epidemiol Biomarkers Prev,2006,15(4):762-768.
33.Hishida A,Matsuo K,Hamajima N,et al.Polymorphism in the hMSH2 gene gIVS126T＞C and risk of non-Hodgkin lymphoma in a Japanese population[J].Cancer Genet Cytogenet,2003,147(1):71-74.
34.Hudler P,Vouk K,Liovic M,et al.Mutations in the hMLH1 gene in Slovenian patients with gastric carcinoma[J].Clin Genet,2004,65(5):405-411.
35.Bevilacqua RA,Simpson AJ.Methylation of the hMLH1 promoter.but no hMLH1mutations in sporadic gastric carcinomas with high-level microsatellite instability[J].Int J Cancer,2000,87(2):200-203.
36.房殿春,罗天辉,李小安,等.胃癌错配修复基因突变和启动子甲基化与基冈不稳定的关系[J].中华消化杂志,2002,22(7):327-330.
37.Menoyo A,Alazzouzi H,Espin E,et al.Smoatic mutations in the DNA damage response genes ATR and CHKI in sporatic stomach tumors with microsatellite instability[J].Cancer Res,2001,61:7727-7730.
38. Yangisawa Y, Akiyama Y, Iida S, et al. Methylation of the MLH1 promoter in familial gastric cancer with microsatellite instability [J]. Int J Cancer, 2000, 85(1):50-53.
39. Kang GH, Shim YH, RO JY. Correlation of methylation of the hMLH1 promoter with lack expression of hMLH1 in sporadic gastric carcinomas with replication error [J]. Lab Invest, 1999,79:903-909.
40. Beak MJ, Kang H, Kim SE, et al. Expression of hMLHl is inactivated in the gastric adenomas with enhanced microsatellite instability [J]. British Journal of Cancer, 2001, 85(8): 1147-1152.
41. Kuismanen SA, Holmberg MT, Salovaara R, et al. Genetic and epigenetic modification of MLH1 accounts for a major share of microsatellite-unstable colorectal cancer [J]. Am. J. Pathol,2000, 156:1773-1779.
42. Nan HM, Song YJ, Yun HY, et al. Effects of dietary intakend genetic factors on hyper methylation of the hMLHl gene promoter in gastric cancer [J]. World J Gastroenterol, 2005, 11(25):3834-3841.
43. Nakajima T, Akiyama Y, Shiraishi J, et al. Age-related hypermethylation of the hMLHl promoter in gastric cancer [J]. Int J Cancer, 2001, 94(2):208-211.
44. Gryfe R, Kim H , Hsieh E T, et al. Tumor microsatellite instability and clinical outcome in young patients with colorectal cancer [J] . N Engl J Med, 2000, 342 (2) : 69-77.
45. Dolcetti R, Viel A, Doglioin C, et al. High prevalence of activa2ted intraepithelial cytotoxic T lymphocytes and increased neoplastic cell apoptosis in colorectal carcinomas with microsatellite instability [J]. A m J Pathol, 1999, 154 (6):1805-1813.
46. Konishi M, Kikuchi Yanoshita R, Tanaka K, et al . Molecular nature of colon tumors in hereditary nonpolyposis colon cancer , familial polyposis , and sporadic colon cancer [J]. Gast roenterology, 1996, 111 (2):307-317.
47. Loeb LA, et al. Multiple mutations and cancer [J]. Proc Natl Acad Sci USA, 2003, 100:776 -81.