IL-1β及其基因多态性在胃癌发生、发展中的作用及机制研究
|
摘要
胃癌是威胁人类健康的主要疾病，在恶性肿瘤中居第二位。环境因素幽门螺杆菌涉及无症状性胃炎到严重的消化性溃疡、甚至胃癌的发生。幽门螺杆菌的致癌作用尚不清楚。而宿主因素如影响IL-1β生成的IL-1B基因多态性亦可决定有的幽门螺杆菌感染患者发展成胃癌，而有的患者则患其它胃病。IL-1β是强烈的胃酸抑制剂，并可影响幽门螺杆菌在胃内的分布。O_2~-·的生成涉及幽门螺杆菌后胃癌的发生与发展。Survivin是凋亡抑制家族的一个新成员，涉及细胞的凋亡和分裂过程。我们研究的目的就是①了解IL-1β、O_2~-·和survivin三者在胃癌中的关系；②Hp诱导人胃粘膜上皮细胞系GES-1细胞IL-1β分泌的细胞内信号转导途径；IL-1β在氧介导胃粘膜细胞增殖中的作用；③IL-1β与胃粘膜上皮细胞survivin表达的关系、靶向survivin对胃癌细胞凋亡的影响；④IL-1B基因(-31、-511)多态性在胃癌、十二指肠溃疡中的作用及与胃癌临床病理参数的关系。旨在探寻IL-1β、O_2~-·和survivin在胃癌中的作用及胃癌治疗的新途径。
IL一lp及其基因多态性在胃癌发生、发展中的作用及机制研究
中文摘要
Hp生长曲线及02’生成情况。观察PMA、DPI、不同浓度SOD对GES一1生成q万
和GES一1增殖的影响。结果:CagA+Hp显著增加GES一1细胞IL一lp的分泌。蛋白
激酶A、C、G抑制剂不能阻断Hp增加的GES一1细胞IL一lp的分泌，蛋白酪氨酸
激酶的抑制剂可阻断Hp增加的GES一1细胞IL一lp的分泌。IL一lp以剂量依赖性增加
GEs一1细胞DNA合成及细胞数量。IL一lp对GES一1细胞的增殖作用可被IL一lra阻断，
IL一lp抗体对此无抑制作用。IL一lp增加GES一1细胞产生02万。GES一1细胞能够自身
产生02丁，PMA增加GES一1细胞产生02万，促进GES一1细胞的增殖。DPI和SOD
减少GEs一1细胞产生02丁，抑制GEs一1细胞的增殖。HP生长情况与接种的菌量有
关，HP能够自身产生大量02丁，其耐受的q丁浓度明显高于胃粘膜细胞。结论:
cagA+HP显著增加GES一1细胞IL一lp的分泌并且依赖于蛋白酪氨酸激酶的活性。
IL一lp刺激可引起GEs一1细胞的增殖及增加GES一1细胞产生02丁。低浓度范围内提
高GEs一1细胞02二生成，可促进OEs一1细胞的增殖.HP能自生。2万，且其耐受02万
的能力高于胃粘膜细胞。IL一lp在调节GEs一1细胞增殖中有重要作用。IL一lp涉及
HP感染胃粘膜细胞的高增殖反应和胃癌的致癌过程。IL一lp促O:丁生成作用可能在
胃相关疾病的防治中具有一定的意义。
Gastric cancer is a major heath problem and remains the second most common cancer in the world. The environment factor, Helicobacter pylori infection is associated with divergent clinical outcomes that from simple asymptomatic gastritis to more serious conditions such as peptic ulcer disease and gastric cancer. The mechanism of H.pylori induced carcinogenesis is not clear. The host genetic factors, such as the polymorphism of IL-1B genes that affects IL-1 production may determine why some individuals infected
with H.pylori develop gastric cancer while others do not. IL-1 is a powerful inhibitor of gastric acid secretion, and can determine the distribution of H.pylori in stomach. The O2
production associated with H.pylori is also involved in the origin and development of gastric cancer. Survivin, a member of the inhibitors of apoptosis protein(IAP) family, is
involved in regulation of apoptosis and cell division. Our study is to make clear ﹖he connection of IL, (V and survivin in gastric cancer; (2)the cellular signal pathway of
CagA+H.pylori inducing IL-1 P secretion in human gastric epithelial (GES-l)cells; the effect of low concentration (V on the proliferation of GES-1 cells. (3)the correlation
between IL and the exprssion of survivin in gastric epithelial cells, the effect of targeting survivin on the apoptosis of gastric cancer cells; (Dthe correlation between polymorphism of IL-1B genes and risk of gastric cancer and duodenal ulcer, the relationship between polymorphism of IL-1B genes and cliniaopathological factors in
human gastric cancer. The final goal of this study is to find the possible role of IL-, O2 ?and survivin in gastric Cancer and supply new strategics in treatment of gastric cancer.
Part 1 The cellular signal pathway of CagA+H. pylori inducing IL-1 protein secretion in GES-1 cells and the effect of IL-1 & on proliferation of GES-1 cells induced by superoxide anion in culture
[Abstract] Objective H. pylori is the main risk factor for the development of gastric cancer. Increased proliferation of the gastric mucosa is a feature of H. pylori infection. The effect of CagA+H.pylori induceing IL-1 P protein secretion in GES-1 cells and various kinase inhibitors on CagA+H. pylori inducing IL-1 P protein secretion, and the effect of IL-lp on gastric epithelial cells proliferation induced by IL-1 Preceptor and superoxide anion in culture have been examined in this study. Metheds GES-1 cells were cultured with CagA+H.pylori in vitro. At the end of culture, IL-1 P protein secretion was assayed by ELISA kit. The effects of the inhibitors of protein kinase A,C,G ,protein tyrosine kinase were analyzed on IL-1 P protein secretion hi GES-1 cells by H.pylori stimulation. GES-1 cells were cultured with IL-1J3. DNA synthesis of GES-1 cells was
assessed by [3H]thymidine incorporation liquid scintillation counter and total viable cell numbers were analyzed by MTT methed. The concentration of 62 ?generated by GES-1
cells and Hp were measured by Ferricytochrome C reduction. Results IL-1 P protein secretion induced by CagA+H.pylori in GES-1 cells was higher than that hi GES-1 cells of never stimulated. Further studied with GES-1 cells showed that IL-1 P protein secretion induced by CagA+ Hp was blocked by the PTK inhibitor (herbimycin A) but not by PKA inhibitor (H7), PKC inhibitor (Calphostin C) and PKG inhibitor (KT5823). IL-lp dose dependency increased DNA synthesis and cell numbers of GES-1 cells. The enhanced
proliferation was blocked by IL-1 receptor antagonist, anti-IL-lp had no this action. Hp can generate CV spontaneously, and endure higher Oa ?concentration than GES-1 cells.
GES-1 cells can also generate (V spontaneously. IL-lp promoted the proliferation of GES-1 cells by stimulating the generation of Ch . Proliferation of GES-1 cells was induced obviously by the physical concentration of C>2 . PMA promoted the proliferation of GES-1 cells by stimulating the generation of C>2 . DPL SOD inhibited the proliferation of GES-1 cells by decreasing the generation of O2 ?Conclusion CagA+ H.pylori
s
IL一lp及其基因多态性在胃癌发生、发展中的作用及机制研究
中文摘要
Hp生长曲线及02’生成情况。观察PMA、DPI、不同浓度SOD对GES一1生成q万
和GES一1增殖的影响。结果:CagA+Hp显著增加GES一1细胞IL一lp的分泌。蛋白
激酶A、C、G抑制剂不能阻断Hp增加的GES一1细胞IL一lp的分泌，蛋白酪氨酸
激酶的抑制剂可阻断Hp增加的GES一1细胞IL一lp的分泌。IL一lp以剂量依赖性增加
GEs一1细胞DNA合成及细胞数量。IL一lp对GES一1细胞的增殖作用可被IL一lra阻断，
IL一lp抗体对此无抑制作用。IL一lp增加GES一1细胞产生02万。GES一1细胞能够自身
产生02丁，PMA增加GES一1细胞产生02万，促进GES一1细胞的增殖。DPI和SOD
减少GEs一1细胞产生02丁，抑制GEs一1细胞的增殖。HP生长情况与接种的菌量有
关，HP能够自身产生大量02丁，其耐受的q丁浓度明显高于胃粘膜细胞。结论:
cagA+HP显著增加GES一1细胞IL一lp的分泌并且依赖于蛋白酪氨酸激酶的活性。
IL一lp刺激可引起GEs一1细胞的增殖及增加GES一1细胞产生02丁。低浓度范围内提
高GEs一1细胞02二生成，可促进OEs一1细胞的增殖.HP能自生。2万，且其耐受02万
的能力高于胃粘膜细胞。IL一lp在调节GEs一1细胞增殖中有重要作用。IL一lp涉及
HP感染胃粘膜细胞的高增殖反应和胃癌的致癌过程。IL一lp促O:丁生成作用可能在
胃相关疾病的防治中具有一定的意义。
Gastric cancer is a major heath problem and remains the second most common cancer in the world. The environment factor, Helicobacter pylori infection is associated with divergent clinical outcomes that from simple asymptomatic gastritis to more serious conditions such as peptic ulcer disease and gastric cancer. The mechanism of H.pylori induced carcinogenesis is not clear. The host genetic factors, such as the polymorphism of IL-1B genes that affects IL-1 production may determine why some individuals infected
with H.pylori develop gastric cancer while others do not. IL-1 is a powerful inhibitor of gastric acid secretion, and can determine the distribution of H.pylori in stomach. The O2
production associated with H.pylori is also involved in the origin and development of gastric cancer. Survivin, a member of the inhibitors of apoptosis protein(IAP) family, is
involved in regulation of apoptosis and cell division. Our study is to make clear ﹖he connection of IL, (V and survivin in gastric cancer; (2)the cellular signal pathway of
CagA+H.pylori inducing IL-1 P secretion in human gastric epithelial (GES-l)cells; the effect of low concentration (V on the proliferation of GES-1 cells. (3)the correlation
between IL and the exprssion of survivin in gastric epithelial cells, the effect of targeting survivin on the apoptosis of gastric cancer cells; (Dthe correlation between polymorphism of IL-1B genes and risk of gastric cancer and duodenal ulcer, the relationship between polymorphism of IL-1B genes and cliniaopathological factors in
human gastric cancer. The final goal of this study is to find the possible role of IL-, O2 ?and survivin in gastric Cancer and supply new strategics in treatment of gastric cancer.
Part 1 The cellular signal pathway of CagA+H. pylori inducing IL-1 protein secretion in GES-1 cells and the effect of IL-1 & on proliferation of GES-1 cells induced by superoxide anion in culture
[Abstract] Objective H. pylori is the main risk factor for the development of gastric cancer. Increased proliferation of the gastric mucosa is a feature of H. pylori infection. The effect of CagA+H.pylori induceing IL-1 P protein secretion in GES-1 cells and various kinase inhibitors on CagA+H. pylori inducing IL-1 P protein secretion, and the effect of IL-lp on gastric epithelial cells proliferation induced by IL-1 Preceptor and superoxide anion in culture have been examined in this study. Metheds GES-1 cells were cultured with CagA+H.pylori in vitro. At the end of culture, IL-1 P protein secretion was assayed by ELISA kit. The effects of the inhibitors of protein kinase A,C,G ,protein tyrosine kinase were analyzed on IL-1 P protein secretion hi GES-1 cells by H.pylori stimulation. GES-1 cells were cultured with IL-1J3. DNA synthesis of GES-1 cells was
assessed by [3H]thymidine incorporation liquid scintillation counter and total viable cell numbers were analyzed by MTT methed. The concentration of 62 ?generated by GES-1
cells and Hp were measured by Ferricytochrome C reduction. Results IL-1 P protein secretion induced by CagA+H.pylori in GES-1 cells was higher than that hi GES-1 cells of never stimulated. Further studied with GES-1 cells showed that IL-1 P protein secretion induced by CagA+ Hp was blocked by the PTK inhibitor (herbimycin A) but not by PKA inhibitor (H7), PKC inhibitor (Calphostin C) and PKG inhibitor (KT5823). IL-lp dose dependency increased DNA synthesis and cell numbers of GES-1 cells. The enhanced
proliferation was blocked by IL-1 receptor antagonist, anti-IL-lp had no this action. Hp can generate CV spontaneously, and endure higher Oa ?concentration than GES-1 cells.
GES-1 cells can also generate (V spontaneously. IL-lp promoted the proliferation of GES-1 cells by stimulating the generation of Ch . Proliferation of GES-1 cells was induced obviously by the physical concentration of C>2 . PMA promoted the proliferation of GES-1 cells by stimulating the generation of C>2 . DPL SOD inhibited the proliferation of GES-1 cells by decreasing the generation of O2 ?Conclusion CagA+ H.pylori
s
引文
1. Wilkinson M: Hclicobacter pylori: An overview. Br J Biomcd Sci 2001; 58: 59-60.
2. Blaser M J, Berg DE: Helicobacter pylori genetic diversity and risk of human disease. J Clin Invest 2001; 107: 767-773.
3．李明峰，凌贞，马爱英等。幽门螺杆菌尿素酶B基因的克隆、表达及免疫源性研究。世界华人消化杂志 1999；7：596-600．
4. Wallace JL. Possible mechanisms and mediators of gastritis associated with Helicobacter pylori infection. Scand J Gastroenterol 1991; 26: 65-70.
5. Godwin CS, Armstrong JA, Marshall BJ. Campylobacter pyloridis, gastritis, and peptic ulceration. J Clin Pathol 1986; 39: 353-365.
6. Cover TL, Blaser MJ. Helicobacter pylori and gastroduodenal disease. Ann Rev Med
1992; 43: 135-145.
7. Blaser MJ. Hypothesis on pathogenesis and natural history of Helicobacter pylori-induced inflammation. Gastroenterology 1992; 102: 720-727.
8. Carrick J, lee A, Hazell S, et al. Campylobaeter pylori, duodenal ulcer, and gastric metaplasia: possible role of functional heterotopie tissues in ulcerogenesis. Gut 1989; 30: 790-797.
9. Megraud F, Lamouliatte H. Helicobacter pylori and duodenal ulcer: evidence suggesting causation. Dig Dis Sci 1992;37: 769-772.
10. Graham DY. Helicobacter pylori: its epidemiology and its role in duodenal ulcer disease. J Gastroenterol Hepathol 1991; 6: 97-105.
11. Huang IQ, Sridhar S, Chen Y, et al. Meta-analysis of the relationship between helicobacter pylori seropositivity and gastric cancer. Gastroenterology 1998; 114: 1169-1179.
12. Danesh I. Helicobacter pylori infection and gastric cancer. Aliment Pharmacol Ther 1999; 13: 851-856.
13. International Agency for Research on Cancer, World Health Organization Schistosomes,. liver flukes and Helicobacter pylori. IARC working group on. the evaluation of carcinogenic risks to human. Monogr Eval Carcinog. Risks Hum 1994; 61: 218-220.
14. Yamaoka Y, Kita M, Kodama T, et al. Induction of various cytokines and development of severe mucosal inflammation by cagA gene positive Helicobacter pylori strains. Gut 1997; 41: 442-451.
15. Wolfe MM, Nompleggi DJ. Cytokine inhibition of gastric acid secretion—a little goes a long way. Gastroenterology 1992; 102:2177-2178.
16. Dinarello CA. Biologic basis for interleukin-1 in disease. Blood 1996; 87: 2095-2147.
17. Robert A, Olafsson AS, Lancaster C, et al. Interleukin-1 is eytoprotective, antisecretory, stimulates PGE2 synthesis by the stomach, and retards gastric emptying. Life Sci 1991; 48: 123-134.
18. Schepp W, Dehne K, Herrmuth H, et al. Identification and functional importance of IL-1 receptors on rat parietal cells. Am J Physiol 1998;275:G1094-1105.
19. Beales IL, Calam J. Interleukin 1 beta and tumour necrosis factor alpha inhibit acid secretion in cultured rabbit parietal cells by multiple pathways. Gut 1998; 42: 227-234.
20. Beales IL, Calam J. Inhibition of carbachol stimulated acid secretion by interleukin-1 β in rabbit parietal cells requires protein kinase C.Gut 2001; 48: 782-789.
21. Prinz C, Neumayer N, Mahr S, et al. Functional impairment of rat enterochromaffin-like cells by interleukin 1 beta. Gastroenterology 1997;112: 364-375.
22. Suzuki K, Grand E, Bowman C et al. TNF-α and interleukin-1 activate gastrin gene expression via MAPK- and PKC-dependent mechanisms Am J Phyiol Gastrointest Liver Physiol 2001 ;281 (6):G1405-1412.
23. Miller LC, Isa S, Vannier E, et al. Live Borrelia burgdorferi preferentially activate interleukin-1 beta gene expression and protein synthesis over the interleukin-1 receptor antagonist. J Clin Invest 1992;90:906-912.
24. Jung HC, Kim JM, Song IS, et al. Helicobacter pylori induces an array of proinflammatory cytokines in human gastric epithelial cells: quantification of mRNA for interleukin-1 alpha/beta, granulocyte-macrophage colonystimulating factor, monocyte chemoattractant protein-1 and tumour necrosis factor-alpha. J Gastroenterol Hepatol 1997; 12:473-480.
25. Basso D, Scrigner M, Toma A, et al. Helicobacter pylori infection enhances mucosal interleukin-1 beta, interleukin-6, and the soluble receptor of interleukin-2. Int J Clin Lab Res 1996;26:207-210.
26. Noach LA, Bosma NB, Jansen J, et al. Mucosal tumor Necro-sisfactor-alpha, interleukin-1beta, andinterleuldn-8 production in patients with Helicobacter pylor iinfection. Scan J Gastroenterology, 1994, 29(5): 425～427.
27. Sharma SA, Tumuru MK, Miller GG, et al. Interleukin-8 response of gastric epithelial cell lines to Helicobacter pylori stimulation in vitro. InfectImmun, 1995; 63(5): 1681-1683
28. Crabtree JE, Shallcross TM, Heatley RV, and Wyatt JI. Mucosal tumour necrosis factor alpha and interleukin-6 in patients with Helicobacter pylori associated gastritis. Gut 1991;32: 1473-1477.
29. El-Omar E, Penman I, Dorrian CA, Ardill JE, and McColl KE. Eradicating Helicobacter pylori infection lowers gastrin mediated acid secretion by two thirds in patients with duodenal ulcer. Gut 1993;34:1060-1065.
30. Graham DY, Opekun A, Lew GM, Evans DJ Jr, Klein PD, and Evans DG. Ablation of exaggerated meal-stimulated gastrin release in duodenal ulcer patients after clearance of Helicobacter(Campylobacter) pylori infection. Am J Gastroenterol 1990;85:394-398.
31. Wang M, Furuta T, Takashima M, et al. Relation between interleukin-lbeta messenger RNA in gastric fundic mucosa and gastric juice pH in patients infected with Helicobacter pylori. J Gastroenterol 1999; 34(suppl 11): 10-17.
32. Takashima M, Furuta T, Hanai H, et al. Efects of Helicobacter pylod infection on gastric acid secretion and serum gastrin levels in Mongolian gerbils. Gut 2001;
41:765-773.
33. Watanabe T, Arakawa T, Fukuda T, et al. Role of neutrophils in a rat model gastric ulcer recurrence cause by IL-1β. Am J Pathol 1997;150:971-979.
34. Hogan DL, Rapier RC, Dreilinger A, et al. Duodenal bicarbonate secretion: eradication of Helicobacter pylori and duodenal structure and function in humans. Gastroenterology 1996; 110:705-716.
35. El-Omar EM, Penman ID, Ardill JE, et al. Helicobacter pylori infection and abnormalities of acid secretion in patients with duodenal ulcer disease. Gastroenterology 1995; 109:681-691.
36. Hannum CH, Wilcox CJ, Arend WP, et al. Interleukin-1 antagonist activity of human interleukin-1 inhibitor. Nature 1990; 343:33-40.
37. Ohtsuki T, Ruetzlar CA, Tasaki K, et al. Interleukin-1 mediates induction of tolerance to global ischemia in gerbil hippocampel CA1 neurons. J Cereb Blood Flow Metab 1996; 16: 1137-1142.
38. Watanabe T, Tada M, Nagai H, et al. Helicobacter pylori infection induces gastric cancer in Mongolian gerbils. Gastroenterology 1998; 115:642-648.
39. El-Omar EM, Carrington M, Chow WH, et al. Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature 2000;404:398-402.
40. Machado JC, Pharoah P, Sousa S, et al. Intedeukin-1B and interleukin-1RN polymorphisms are associated with increased risk of gastric carcinoma. Gastroenterology 2001; 121:823-829.
41. Takahisa F, EL-Omar EM, Fang X, et al. Interleukin-1B polymorphisms increase risk of hypochlorhydria and atrophic gastric and reduced risk of duodenal ulcer recurrence in Japan. Gastroenterology 2002; 123:92-105.
42. Furuta T, Shirai N, Xiao F, et al. Effect of genotypic differences in interleukin-1beta on gastric acid secretion, gastric artrophy, and recurrence of peptic deer disease. Gastroenterology 2000;120(suppl 1):A649.
43. EL-Omar EM, Chow WH, Rabin CS. Gastric cancer and H.pylori: host genetics open the way. Gastroenterology 2001; 121:1002-1013.
44. Shigetada T, Kazuhito R, Takeshi N, et al. Guinea pig gastric mucosal cells produce abundant superoxide anion through an NADPH oxidase-like system. Gastroenterology 1998; 115:1186-1196.
45. Mikhalchik EV, Kharaeva ZF, Kovalchuk LV, et al. Effect of cytokins on the level of free radical in the blood of patients with systemic and local staphylococcus infection. Russ J Immunol 2002; 7(3):251-258.
46. Sakai H, Ohira Y, Sunaka A, et al. Inhibition of small-conductance C1- channels by the interleukin-1 beta-stimulated production of superoxide in rabbit gastric parietal
cells. J physiol 2003 ;511 (15):207-217.
47. Kelly A, Lynch A, Vereker E, et al. The Anti-inflammatory Cytokine, Interleukin (IL)-10, Blocks the Inhibitory Effect of IL-1β on Long Term Potentiation. J of Bio Chem 2001; 276(49): 45564-45572.
48. Phull PS, Green CJ, Jacyna MR. A radical view of the stomach: The role of oxygen-derived free radicals and oxidants in gastrodudenal disease. Eur J Gastroenterol Hepatol. 1995:7:265-274.
49. Davies GR, Simmonds NJ, Stevens TRJ, et al. Helicobacter pylori stimulates antral mucosal reactive oxygen species. Gut 1994: 35:179-185.
50. Brown LM. Helicobacter pylori: Epidemiology and routes of transmission. Epidemiol Rev 2000; 22:283-297.
51. Graham S, Haughey B, Marshall J, et al. Diet in the epidemiology of gastric cancer. Nutr. Cancer 1990;13: 19-34.
52. La Vecchia C, Decarli A, Negri E. and Parazzini F. Epidemiological aspects of diet and cancer: a summary review of case-control studies from northern Italy. Ontology 1988; 45: 364-370.
53. Suzuki M, Nakamura M, Mori M, et al. Lansoprazole inhibits oxygen-derived free radical production from neutrophils activated by Helicobacter pylori. J. Clin. Gastroenterol 1995; 20 (Suppl. 2): S93-S96.
54. Davies G R, Banatvala N, Collins C E,et al. Relationship between infective load of Helicobacter pylori and reactive oxygen metabolite production in antral mucosa. Stand. J. Gastroenterol 1994; 29: 419-424.
55. Yoshida N, Yoshikawa T, Iinuma S, et al. Rebamipide protects against activation of neutrophils by Helicobacter pylori. Digest. Dis. Sci., 1996; 41:1139-1144.
56. Farinati F, Cardin R, Degan P, et al. Oxidative DNA damage accumulation in gastric carcinogenesis. Gut 1998; 42(3):351-356.
57. Bagchi D, Mcginn TR, Ye X, et al. Helicobacter pylori-induced oxidative stress and DNA damage in a primary culture of human gastric mucosal cells. Dig Dis Sci 2002;47(6): 1405-1412.
58. Nicotera P, Kass GEN, Duddy DK, et al. Calcium and Signal transduction in oxidative cell damage. In: Duncan CJ, eds. Alcium, oxygen radicals and cellular damage. Cambridge: Cambridge University Press 1991: 17-33.
59. Nathan CF, Silverstein SC, Brukner LH, et al. Extracellular cytolysis by activared macrophages and granulocytes.Ⅱ. Hydrogen peroxide as a mediator of cytotoxicity. J Exp Med 1979; 149: 100-113.
60. Babior BM. Oxidants form phagocytes: agents of defense and destruction. Blood 1984; 64: 959-966.
61. Weiss SJ, Lobuglio AF. Phagocyte-generated oxygen metabolities and cellular injury. Lab Invest 1982; 47:5-18.
62. Yoshida N, Granger DN, Evans DJ Jr, et al. Mechanism involved in Helicobacter pylori-induced inflammation. Gastroenterology 1993; 105:1431-1440.
63. Bagchi D, Bhattacharya G, Stohs SJ. Production of reactive oxygen species by gastric cells in association with Helicobacter pylori. Free Radic Res 1996;24:439-450.
64. Teshima S, Kutsumi H, Kawahara T, et al. Regulation of growth and apoptosis of cultured guinea pig gastric mucosal cells by mitogenic oxidase 1. Am J Physiol Gastrointest Liver Physiol 2000; 279:G1169-G1176.
65. Ames BN, Gold LS. Too many rodent carcinogens: Mitogenesis increase mutagenesis. Science 1991;249:970.
66. Parsonnet J. Helicobacter pylori and gastric cancer. Gastroenterol Clin of North America 1993;22:89-104.
67. Pervaiz S, Clement MV. A permissive apoptotic environment: function of a decrease in intercellular superoxide anion and cytosolic acidification. Biolchemical and Biophysical Research Communications 2002; 290:1145-1150.
68. Dreher D, Junod AF. Role of oxygen free radicals in cancer development. Eur J Cancer 1996;32A: 30-38.
69. Adida C, Crotty PL, McGrath J, et al. Developmentally regulated expression of the novel cancer antiapoptosis gene survivin in human and mouse differentiation.Am J Pathol 1998; 152(1):43-49.
70. Verhaegen S, Adrian J, McGovan J, et al. Inhibition of apoptosis by antioxidants in the human HL-60 leukemia cell line. Biochem Pharmacol 1995 ;40:1021-1029.
71. Konno R, Yamakawa H, Utsunomiya H, et al. Expression of survivin and bcl-2 in the normal human endometrium.Mol Hum Reprod 2000; 6(6):529-534.
72. Kawasaki H, Altieri DC, Lu CD, et al. Inhibition of apoptosis by survivn predicts rats in colorectal cancer. Cancer Res 1998,58(22):5071-5074.
73. Johnstone RW, Ruefli AA, Lowe SW. Apoptosis: a link between cancer genetics and chemotherapy. Cell 2002,108:153-164.
74. Ambrosini G, Adida C, Altieri DC, et al. A novel antiapoptosis gene, survivin expressed in cancer and lymphoma. Nat Med 1997;3(8):917-921.
75. O'Cormor DS, Grossman D, Plescia J, et al. Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. Proc Nat Acad Sci USA 2000,97:13103-13107.
76. Verdecia MA, Huang HK, Dutil E, et al. Structure of the human anti-apoptotic protein survivin reveals a dimeric arrangement. Nat Struct Biol 2000;7(7):602-608.
77. Tamm I, Wang Y, Sausville E, et al. IAP family protein survivin inhibits caspase activity and apoptosis induced by Fas(CD95), Bax, caspases,and anticancer drugs. Cancer Res 1998;58(23):5315-5320.
78. Li F, Ambrosini G, Chu EY, et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 1998;396(6711):580-584.
79. Yamamoto S, Yasui W, Kitadai Y, et al. Expression of vascular endothelial growth factor in human gastric carcinomas. Pathol Int 1998;48(7):499-506.
80. Lu CD, Altieri DC, Tanigawa N. Expression of a novel antiapoptosis gene: survivin, correlated with tumor cell apoptosis and P53 accumulation in gastric carcinomas.Cancer Res 1998 ;58(9): 1808-1812.
81. kasagi N, Gomyo Y, Shirai H, et al. Apoptotic cell death in human gastric carcinoma: analysis by terminal deoxynucleotidyl transforase-mediated dUTP-biotin nick end labeling. Jpn J Cancer Res 1994,85(9):939-948.
82. Grossman D, Kim PJ, Blanc-Brude OP, et al. transgenic expression of survivin in keratinocytes counteracts UVB induced apoptosis and cooperate with loss of P53. J Clin Invest 2001,108:991-999.
83. Okada E, Murai Y, Matsui K, et al. Survivin expression in tumor cell nuclei is predictive of a favorable prognosis in gastric cancer patients. Cancer Lett 2001,163:109-116.
84. Lanwera GY, Scoot GV, Karpeh MS, et al. Immunohistochemical evaluation of bcl-2 protein expression in gastric adenocarcinoma. Cancer 1995; 75:2209
85. Buolawini JK. Novel anticancer drug discovery. Curr Opin Chem Biol 1999;3(4):500-509.
86. Tu SP, Jiang XH, Lin MCM, et al. Suppression of survivin expression inhibits in vivo tumorigenicity and angiogenesis in gastric cancer. Cancer Res 2003;65(15): 7724-7732.
87．孙璟，涂水平，谭继宏等．生存素基因突变体诱导胃癌细胞有丝分裂障碍的实验研究．中华消化杂志 2003；23 (8)：487-490．
88．谭继宏，涂水平，邹冰等．生存素基因突变体诱导胃癌细胞凋亡的实验研究．中华消化杂志 2003；23 (4)：199-202．
89. Olie RA, Simoes-Wust AP, Baumann B, et al. a novel antisense oligonucleotide targeting survivn expewssion induces apoptosis and ensitizes lung cancer cells to chemotherapy. Cancer Res 2000;60(11):2805-2809.
90．李国华，杨建辉．Survivin基因反义寡核苷酸增强胃癌细胞对顺铂的敏感性研究．肿瘤 2003；23 (4)：297-299．
2. Blaser M J, Berg DE: Helicobacter pylori genetic diversity and risk of human disease. J Clin Invest 2001; 107: 767-773.
3．李明峰，凌贞，马爱英等。幽门螺杆菌尿素酶B基因的克隆、表达及免疫源性研究。世界华人消化杂志 1999；7：596-600．
4. Wallace JL. Possible mechanisms and mediators of gastritis associated with Helicobacter pylori infection. Scand J Gastroenterol 1991; 26: 65-70.
5. Godwin CS, Armstrong JA, Marshall BJ. Campylobacter pyloridis, gastritis, and peptic ulceration. J Clin Pathol 1986; 39: 353-365.
6. Cover TL, Blaser MJ. Helicobacter pylori and gastroduodenal disease. Ann Rev Med
1992; 43: 135-145.
7. Blaser MJ. Hypothesis on pathogenesis and natural history of Helicobacter pylori-induced inflammation. Gastroenterology 1992; 102: 720-727.
8. Carrick J, lee A, Hazell S, et al. Campylobaeter pylori, duodenal ulcer, and gastric metaplasia: possible role of functional heterotopie tissues in ulcerogenesis. Gut 1989; 30: 790-797.
9. Megraud F, Lamouliatte H. Helicobacter pylori and duodenal ulcer: evidence suggesting causation. Dig Dis Sci 1992;37: 769-772.
10. Graham DY. Helicobacter pylori: its epidemiology and its role in duodenal ulcer disease. J Gastroenterol Hepathol 1991; 6: 97-105.
11. Huang IQ, Sridhar S, Chen Y, et al. Meta-analysis of the relationship between helicobacter pylori seropositivity and gastric cancer. Gastroenterology 1998; 114: 1169-1179.
12. Danesh I. Helicobacter pylori infection and gastric cancer. Aliment Pharmacol Ther 1999; 13: 851-856.
13. International Agency for Research on Cancer, World Health Organization Schistosomes,. liver flukes and Helicobacter pylori. IARC working group on. the evaluation of carcinogenic risks to human. Monogr Eval Carcinog. Risks Hum 1994; 61: 218-220.
14. Yamaoka Y, Kita M, Kodama T, et al. Induction of various cytokines and development of severe mucosal inflammation by cagA gene positive Helicobacter pylori strains. Gut 1997; 41: 442-451.
15. Wolfe MM, Nompleggi DJ. Cytokine inhibition of gastric acid secretion—a little goes a long way. Gastroenterology 1992; 102:2177-2178.
16. Dinarello CA. Biologic basis for interleukin-1 in disease. Blood 1996; 87: 2095-2147.
17. Robert A, Olafsson AS, Lancaster C, et al. Interleukin-1 is eytoprotective, antisecretory, stimulates PGE2 synthesis by the stomach, and retards gastric emptying. Life Sci 1991; 48: 123-134.
18. Schepp W, Dehne K, Herrmuth H, et al. Identification and functional importance of IL-1 receptors on rat parietal cells. Am J Physiol 1998;275:G1094-1105.
19. Beales IL, Calam J. Interleukin 1 beta and tumour necrosis factor alpha inhibit acid secretion in cultured rabbit parietal cells by multiple pathways. Gut 1998; 42: 227-234.
20. Beales IL, Calam J. Inhibition of carbachol stimulated acid secretion by interleukin-1 β in rabbit parietal cells requires protein kinase C.Gut 2001; 48: 782-789.
21. Prinz C, Neumayer N, Mahr S, et al. Functional impairment of rat enterochromaffin-like cells by interleukin 1 beta. Gastroenterology 1997;112: 364-375.
22. Suzuki K, Grand E, Bowman C et al. TNF-α and interleukin-1 activate gastrin gene expression via MAPK- and PKC-dependent mechanisms Am J Phyiol Gastrointest Liver Physiol 2001 ;281 (6):G1405-1412.
23. Miller LC, Isa S, Vannier E, et al. Live Borrelia burgdorferi preferentially activate interleukin-1 beta gene expression and protein synthesis over the interleukin-1 receptor antagonist. J Clin Invest 1992;90:906-912.
24. Jung HC, Kim JM, Song IS, et al. Helicobacter pylori induces an array of proinflammatory cytokines in human gastric epithelial cells: quantification of mRNA for interleukin-1 alpha/beta, granulocyte-macrophage colonystimulating factor, monocyte chemoattractant protein-1 and tumour necrosis factor-alpha. J Gastroenterol Hepatol 1997; 12:473-480.
25. Basso D, Scrigner M, Toma A, et al. Helicobacter pylori infection enhances mucosal interleukin-1 beta, interleukin-6, and the soluble receptor of interleukin-2. Int J Clin Lab Res 1996;26:207-210.
26. Noach LA, Bosma NB, Jansen J, et al. Mucosal tumor Necro-sisfactor-alpha, interleukin-1beta, andinterleuldn-8 production in patients with Helicobacter pylor iinfection. Scan J Gastroenterology, 1994, 29(5): 425～427.
27. Sharma SA, Tumuru MK, Miller GG, et al. Interleukin-8 response of gastric epithelial cell lines to Helicobacter pylori stimulation in vitro. InfectImmun, 1995; 63(5): 1681-1683
28. Crabtree JE, Shallcross TM, Heatley RV, and Wyatt JI. Mucosal tumour necrosis factor alpha and interleukin-6 in patients with Helicobacter pylori associated gastritis. Gut 1991;32: 1473-1477.
29. El-Omar E, Penman I, Dorrian CA, Ardill JE, and McColl KE. Eradicating Helicobacter pylori infection lowers gastrin mediated acid secretion by two thirds in patients with duodenal ulcer. Gut 1993;34:1060-1065.
30. Graham DY, Opekun A, Lew GM, Evans DJ Jr, Klein PD, and Evans DG. Ablation of exaggerated meal-stimulated gastrin release in duodenal ulcer patients after clearance of Helicobacter(Campylobacter) pylori infection. Am J Gastroenterol 1990;85:394-398.
31. Wang M, Furuta T, Takashima M, et al. Relation between interleukin-lbeta messenger RNA in gastric fundic mucosa and gastric juice pH in patients infected with Helicobacter pylori. J Gastroenterol 1999; 34(suppl 11): 10-17.
32. Takashima M, Furuta T, Hanai H, et al. Efects of Helicobacter pylod infection on gastric acid secretion and serum gastrin levels in Mongolian gerbils. Gut 2001;
41:765-773.
33. Watanabe T, Arakawa T, Fukuda T, et al. Role of neutrophils in a rat model gastric ulcer recurrence cause by IL-1β. Am J Pathol 1997;150:971-979.
34. Hogan DL, Rapier RC, Dreilinger A, et al. Duodenal bicarbonate secretion: eradication of Helicobacter pylori and duodenal structure and function in humans. Gastroenterology 1996; 110:705-716.
35. El-Omar EM, Penman ID, Ardill JE, et al. Helicobacter pylori infection and abnormalities of acid secretion in patients with duodenal ulcer disease. Gastroenterology 1995; 109:681-691.
36. Hannum CH, Wilcox CJ, Arend WP, et al. Interleukin-1 antagonist activity of human interleukin-1 inhibitor. Nature 1990; 343:33-40.
37. Ohtsuki T, Ruetzlar CA, Tasaki K, et al. Interleukin-1 mediates induction of tolerance to global ischemia in gerbil hippocampel CA1 neurons. J Cereb Blood Flow Metab 1996; 16: 1137-1142.
38. Watanabe T, Tada M, Nagai H, et al. Helicobacter pylori infection induces gastric cancer in Mongolian gerbils. Gastroenterology 1998; 115:642-648.
39. El-Omar EM, Carrington M, Chow WH, et al. Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature 2000;404:398-402.
40. Machado JC, Pharoah P, Sousa S, et al. Intedeukin-1B and interleukin-1RN polymorphisms are associated with increased risk of gastric carcinoma. Gastroenterology 2001; 121:823-829.
41. Takahisa F, EL-Omar EM, Fang X, et al. Interleukin-1B polymorphisms increase risk of hypochlorhydria and atrophic gastric and reduced risk of duodenal ulcer recurrence in Japan. Gastroenterology 2002; 123:92-105.
42. Furuta T, Shirai N, Xiao F, et al. Effect of genotypic differences in interleukin-1beta on gastric acid secretion, gastric artrophy, and recurrence of peptic deer disease. Gastroenterology 2000;120(suppl 1):A649.
43. EL-Omar EM, Chow WH, Rabin CS. Gastric cancer and H.pylori: host genetics open the way. Gastroenterology 2001; 121:1002-1013.
44. Shigetada T, Kazuhito R, Takeshi N, et al. Guinea pig gastric mucosal cells produce abundant superoxide anion through an NADPH oxidase-like system. Gastroenterology 1998; 115:1186-1196.
45. Mikhalchik EV, Kharaeva ZF, Kovalchuk LV, et al. Effect of cytokins on the level of free radical in the blood of patients with systemic and local staphylococcus infection. Russ J Immunol 2002; 7(3):251-258.
46. Sakai H, Ohira Y, Sunaka A, et al. Inhibition of small-conductance C1- channels by the interleukin-1 beta-stimulated production of superoxide in rabbit gastric parietal
cells. J physiol 2003 ;511 (15):207-217.
47. Kelly A, Lynch A, Vereker E, et al. The Anti-inflammatory Cytokine, Interleukin (IL)-10, Blocks the Inhibitory Effect of IL-1β on Long Term Potentiation. J of Bio Chem 2001; 276(49): 45564-45572.
48. Phull PS, Green CJ, Jacyna MR. A radical view of the stomach: The role of oxygen-derived free radicals and oxidants in gastrodudenal disease. Eur J Gastroenterol Hepatol. 1995:7:265-274.
49. Davies GR, Simmonds NJ, Stevens TRJ, et al. Helicobacter pylori stimulates antral mucosal reactive oxygen species. Gut 1994: 35:179-185.
50. Brown LM. Helicobacter pylori: Epidemiology and routes of transmission. Epidemiol Rev 2000; 22:283-297.
51. Graham S, Haughey B, Marshall J, et al. Diet in the epidemiology of gastric cancer. Nutr. Cancer 1990;13: 19-34.
52. La Vecchia C, Decarli A, Negri E. and Parazzini F. Epidemiological aspects of diet and cancer: a summary review of case-control studies from northern Italy. Ontology 1988; 45: 364-370.
53. Suzuki M, Nakamura M, Mori M, et al. Lansoprazole inhibits oxygen-derived free radical production from neutrophils activated by Helicobacter pylori. J. Clin. Gastroenterol 1995; 20 (Suppl. 2): S93-S96.
54. Davies G R, Banatvala N, Collins C E,et al. Relationship between infective load of Helicobacter pylori and reactive oxygen metabolite production in antral mucosa. Stand. J. Gastroenterol 1994; 29: 419-424.
55. Yoshida N, Yoshikawa T, Iinuma S, et al. Rebamipide protects against activation of neutrophils by Helicobacter pylori. Digest. Dis. Sci., 1996; 41:1139-1144.
56. Farinati F, Cardin R, Degan P, et al. Oxidative DNA damage accumulation in gastric carcinogenesis. Gut 1998; 42(3):351-356.
57. Bagchi D, Mcginn TR, Ye X, et al. Helicobacter pylori-induced oxidative stress and DNA damage in a primary culture of human gastric mucosal cells. Dig Dis Sci 2002;47(6): 1405-1412.
58. Nicotera P, Kass GEN, Duddy DK, et al. Calcium and Signal transduction in oxidative cell damage. In: Duncan CJ, eds. Alcium, oxygen radicals and cellular damage. Cambridge: Cambridge University Press 1991: 17-33.
59. Nathan CF, Silverstein SC, Brukner LH, et al. Extracellular cytolysis by activared macrophages and granulocytes.Ⅱ. Hydrogen peroxide as a mediator of cytotoxicity. J Exp Med 1979; 149: 100-113.
60. Babior BM. Oxidants form phagocytes: agents of defense and destruction. Blood 1984; 64: 959-966.
61. Weiss SJ, Lobuglio AF. Phagocyte-generated oxygen metabolities and cellular injury. Lab Invest 1982; 47:5-18.
62. Yoshida N, Granger DN, Evans DJ Jr, et al. Mechanism involved in Helicobacter pylori-induced inflammation. Gastroenterology 1993; 105:1431-1440.
63. Bagchi D, Bhattacharya G, Stohs SJ. Production of reactive oxygen species by gastric cells in association with Helicobacter pylori. Free Radic Res 1996;24:439-450.
64. Teshima S, Kutsumi H, Kawahara T, et al. Regulation of growth and apoptosis of cultured guinea pig gastric mucosal cells by mitogenic oxidase 1. Am J Physiol Gastrointest Liver Physiol 2000; 279:G1169-G1176.
65. Ames BN, Gold LS. Too many rodent carcinogens: Mitogenesis increase mutagenesis. Science 1991;249:970.
66. Parsonnet J. Helicobacter pylori and gastric cancer. Gastroenterol Clin of North America 1993;22:89-104.
67. Pervaiz S, Clement MV. A permissive apoptotic environment: function of a decrease in intercellular superoxide anion and cytosolic acidification. Biolchemical and Biophysical Research Communications 2002; 290:1145-1150.
68. Dreher D, Junod AF. Role of oxygen free radicals in cancer development. Eur J Cancer 1996;32A: 30-38.
69. Adida C, Crotty PL, McGrath J, et al. Developmentally regulated expression of the novel cancer antiapoptosis gene survivin in human and mouse differentiation.Am J Pathol 1998; 152(1):43-49.
70. Verhaegen S, Adrian J, McGovan J, et al. Inhibition of apoptosis by antioxidants in the human HL-60 leukemia cell line. Biochem Pharmacol 1995 ;40:1021-1029.
71. Konno R, Yamakawa H, Utsunomiya H, et al. Expression of survivin and bcl-2 in the normal human endometrium.Mol Hum Reprod 2000; 6(6):529-534.
72. Kawasaki H, Altieri DC, Lu CD, et al. Inhibition of apoptosis by survivn predicts rats in colorectal cancer. Cancer Res 1998,58(22):5071-5074.
73. Johnstone RW, Ruefli AA, Lowe SW. Apoptosis: a link between cancer genetics and chemotherapy. Cell 2002,108:153-164.
74. Ambrosini G, Adida C, Altieri DC, et al. A novel antiapoptosis gene, survivin expressed in cancer and lymphoma. Nat Med 1997;3(8):917-921.
75. O'Cormor DS, Grossman D, Plescia J, et al. Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin. Proc Nat Acad Sci USA 2000,97:13103-13107.
76. Verdecia MA, Huang HK, Dutil E, et al. Structure of the human anti-apoptotic protein survivin reveals a dimeric arrangement. Nat Struct Biol 2000;7(7):602-608.
77. Tamm I, Wang Y, Sausville E, et al. IAP family protein survivin inhibits caspase activity and apoptosis induced by Fas(CD95), Bax, caspases,and anticancer drugs. Cancer Res 1998;58(23):5315-5320.
78. Li F, Ambrosini G, Chu EY, et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 1998;396(6711):580-584.
79. Yamamoto S, Yasui W, Kitadai Y, et al. Expression of vascular endothelial growth factor in human gastric carcinomas. Pathol Int 1998;48(7):499-506.
80. Lu CD, Altieri DC, Tanigawa N. Expression of a novel antiapoptosis gene: survivin, correlated with tumor cell apoptosis and P53 accumulation in gastric carcinomas.Cancer Res 1998 ;58(9): 1808-1812.
81. kasagi N, Gomyo Y, Shirai H, et al. Apoptotic cell death in human gastric carcinoma: analysis by terminal deoxynucleotidyl transforase-mediated dUTP-biotin nick end labeling. Jpn J Cancer Res 1994,85(9):939-948.
82. Grossman D, Kim PJ, Blanc-Brude OP, et al. transgenic expression of survivin in keratinocytes counteracts UVB induced apoptosis and cooperate with loss of P53. J Clin Invest 2001,108:991-999.
83. Okada E, Murai Y, Matsui K, et al. Survivin expression in tumor cell nuclei is predictive of a favorable prognosis in gastric cancer patients. Cancer Lett 2001,163:109-116.
84. Lanwera GY, Scoot GV, Karpeh MS, et al. Immunohistochemical evaluation of bcl-2 protein expression in gastric adenocarcinoma. Cancer 1995; 75:2209
85. Buolawini JK. Novel anticancer drug discovery. Curr Opin Chem Biol 1999;3(4):500-509.
86. Tu SP, Jiang XH, Lin MCM, et al. Suppression of survivin expression inhibits in vivo tumorigenicity and angiogenesis in gastric cancer. Cancer Res 2003;65(15): 7724-7732.
87．孙璟，涂水平，谭继宏等．生存素基因突变体诱导胃癌细胞有丝分裂障碍的实验研究．中华消化杂志 2003；23 (8)：487-490．
88．谭继宏，涂水平，邹冰等．生存素基因突变体诱导胃癌细胞凋亡的实验研究．中华消化杂志 2003；23 (4)：199-202．
89. Olie RA, Simoes-Wust AP, Baumann B, et al. a novel antisense oligonucleotide targeting survivn expewssion induces apoptosis and ensitizes lung cancer cells to chemotherapy. Cancer Res 2000;60(11):2805-2809.
90．李国华，杨建辉．Survivin基因反义寡核苷酸增强胃癌细胞对顺铂的敏感性研究．肿瘤 2003；23 (4)：297-299．