間白素-1(interleukin-1，IL-1)是種多功能的細胞間素(cytokine)，此家族包括間白素-1α (interleukin-1α，IL-1α)、間白素-1β (interleukin-1β，IL-1β)和間白素接受器拮抗子(interleukin-1 receptor antagonist，IL-1Ra)。IL-1幾乎是可以影響每種細胞，也可經由許多細胞產生。其中IL-1β是可以抗潰瘍、胃部的細胞內保護劑、拮抗分泌(antisecretory)和胃部排空的抑制劑；IL-1Ra則是與IL-1β競爭間白素接受器(interleukin-1 receptor，IL-1R)。IL-1被發現在高加索人種當中對胃癌有相當的敏感性，亞洲人種也有相同的敏感性。十二指腸潰瘍和胃癌都是由複雜的原因造成的，但其主因都是胃酸分泌。所以我們希望利用間白素-1β和間白素接受器拮抗子的基因多型性(gene polymorphisms)和探討不同檢體的基因型是否與胃癌或是十二指腸潰瘍有關。我們使用了PCR-CTPP、RFLP和PCR-VNTR等實驗方法，測得140個胃癌病患、94個十二指腸潰瘍病患與160個正常人檢體的基因型；最後以統計方法計算出其間之相關性。
結果顯示IL-1RN 2R對偶基因型的胃癌與正常人的比較有顯著差異(9 % vs. 3%, p = 0.781)，而在其他基因位置如IL-1B -31、IL-1B -511和IL-1B +3954並沒有什麼差異。攜帶IL-1RN 2R基因型的胃癌患者發展成小腸型(intestinal type)胃癌的危險性增高，因其危險對比值(odds ratio，OR)為4.06 (95 % CI: 1.68 - 9.79，p-value=0.085)。而攜帶IL-1RN 2R基因型的胃癌患者發展成瀰漫型(diffuse type)胃癌的危險性增高，因其危險對比值(odds ratio，OR)為3.15 (95 % CI: 1.16-8.56，p-value=0.061)有增加其危險性。IL-1RN基因型在十二指腸潰瘍發展中會增加其危險性(OR= 2.57, 95 % CI: 1.03-6.83, p-value= 0.292)，但在IL-1B基因型當中並沒有明顯的不同。除此之外，我們實驗顯示血型為A型同時又攜帶IL-1RN 2R基因型的人會增加罹患胃癌的危險性(OR= 4.51, 95 % CI: 1.20-16.88，p-value=0.516)；血型為O型同時又攜帶IL-1RN 2R基因型的人，也會增加罹患十二指腸潰瘍的危險性(OR= 10.32, 95 % CI: 2.10-50.61，p-value= 0.160)。
結果顯示攜帶IL-1RN 2R對偶基因型的胃癌及十二指腸潰瘍患者之患病機會是IL-RN 4R的3倍以上。受到幽門螺旋桿菌感染、飲酒或是吸菸等外在環境的影響，雖然會提高胃癌和十二指腸潰瘍的危險性，但其主要還是因為攜帶了IL-1RN 2R基因型所造成的。IL-1RN 2R基因型雖然是造成胃癌和十二指腸潰瘍的主要基因型，如果再加上血型的遺傳因素，對於罹患胃癌及十二指腸潰瘍危險性的提高有加成性的相互影響。

Interleukin-1 (IL-1) is a prototypic multifunctional cytokine. IL-1 family include interleukin-1 a (IL-1 a), interleukin-1b (IL-1 b) and interleukin-1 receptor antagonist (IL-1 Ra). IL-1 b is the archetypeal pleiotropic cytokine which have been produced by many cells and exerting its biological effects on almost all cell types. IL-1 b is the most potent of known agents that are gastric cytoprotective, antiulcer, antisecretory and an inhibitor of gastric emptying. IL-1 Ra competes with IL-1 b for cell surface receptor occupancy. Host genetic factors that affect interleukin-1 (IL-1) have been reported to influence the susceptibility of Caucasians to gastric cancer. Whether Asians have the same genetic susceptibility remains unclear. In this study, the genetic associations of IL-1B and IL-1RN polymorphisms with gastric cancer and duodenal ulcer in Taiwan were evaluated.
Genomic DNA from 140 unrelated Taiwanese patients with gastric adenocarcinoma, 94 with duodenal ulcer and 165 ethically matched healthy controls was typed for polymorphisms at positions –31, -511, and +3954 in the IL-1B gene, and the variable number of tandem repeats polymorphisms in intron 2 of the IL-1RN gene.
The allele frequencies of IL-1RN 2R in gastric cancer cases were much higher than those in healthy controls (9% vs. 3%, p = 0.781). The allele frequencies of IL-1B –31, IL-1B –511 and IL-1B +3954 did not differ. An increased risk of the development of intestinal type gastric carcinoma was found in IL-1RN 2R carriers with an odds ratio (OR) of 4.06 (95% confidence interval [CI]: 1.68 – 9.79, p-value=0.085). And another increased risk of the development of diffuse type gastric carcinoma was found in IL-1RN 2R carriers with an odds ratio (OR) of 3.15 (95% confidence interval [CI]: 1.16 – 8.56, p-value=0.061). A significant association was found in IL-1RN 2R/4R genotype and the risk of the development of duodenal ulcer, with an odds ratio (OR) of 2.57 (95% CI: 1.03 – 6.38, p = 0.292). No significant relationship was noted in duodenal ulcer patients with IL-1B genotype examed in this study. Additionally, a synergistic interaction between blood type A and IL-1 RN 2R carriers existed in gastric cancer patients (OR= 4.51; 95% CI: 1.20 – 16.88, p-value=0.516). The synergistic interaction was even stronger between blood type O and IL-1 RN 2R carriers of duodenal ulcer patients (OR= 10.3; 95% CI: 2.10 – 50.61, p-value=0.160).
In conclusion, the genetic polymorphisms of IL-1RN 2R and blood type A are associated with the development of gastric cancer. The genetic polymorphisms of IL-1RN 2R and blood type O are associated with the development of duodenal ulcer.

目錄
中文摘要………………………………………………………… i
英文摘要………………………………………………………… iii
壹､緒論…………………..…………………………………… 1
一､間白素-1………….……………………………………… 1
二､幽門螺旋桿菌…………………………………………… 5
三､十二指腸潰瘍…………………………………………… 7
四､胃癌的發展……………………………………………… 10
(一)胃炎……………………………………………………… 11
(二)胃息肉…………………………………………………… 12
(三)胃癌….……………………………………………… 13
貳､研究目的………………………………………………… 16
參､實驗材料與方法………………………………………… 17
一､實驗材料………………………………………………… 17
二､實驗流程圖……………………………………………… 19
三､實驗方法………………………………………………… 20
(一)血液和組織DNA萃取…………………………………… 20
(二)PCR-CTPP、RLFP和PCR的實驗方法…………………… 22
1.primers的設計…………………………………………… 22
2.PCR-CTPP、RFLP和PCR…………………………………… 24
(1)IL-1B –31 PCR-CTPP………………………………… 24
(2)IL-1B –511 RLFP…………………………………… 26
(3)IL-1B +3954 RLFP…………………………………… 27
(4)IL-1RN PCR…………………………………………… 29
(三)統計方法……………………………………………… 30
肆､結果…………………………………………………… 31
一､胃癌……………………………………………………… 31
二､十二指腸潰瘍…………………………………………… 37
伍､討論……………………………………………………… 42
參考文獻……………………………………………………… 49
表目錄
表一 胃癌病人與正常人在IL-1B and IL-1RN 基因型之比較 55
表二 胃癌病人與正常人在IL-1B and IL-1RN 對偶基因型之
比較………………………………………………………… 56
表三 IL-1B 和 IL-1RN haplotype frequencies 對罹患胃癌危險性之比較……………………………………………………… 57
表四 IL-1B和IL-1RN基因型對於造成腸型(intestinal)及瀰漫型(diffuse)胃癌危險性之比較……………………………………… 58
表五 H.pylori感染與IL-1B和IL-1 RN基因型對於罹患胃癌危險性之比較…………………………………………………… 59
表六 血型 (A型與非A型)與IL-1 B and IL-1 RN基因型對於罹患胃癌危險性之比較……………………………………… 60
表七 吸煙習慣與IL-1B和IL-1RN基因型對於罹患胃癌危險性之比較………………………………………………………… 61
表八 飲酒習慣與IL-1B和IL-1RN基因型對罹患胃癌危險性之比較
………………………………………………………… 62
表九 十二指腸潰瘍病人與正常人在IL-1B and IL-1RN 基因型之比較……………………………………………………… 63
表十 十二指腸潰瘍病人與正常人在IL-1B and IL-1RN 對偶基因型之比較………………………………………………… 64
表十一 IL-1B 和 IL-1RN haplotype frequencies 對罹患十二指腸潰瘍危險性之比較………………………………………… 65
表十二(a) 血型 (O型與非O型)與IL-1 B and IL-1 RN基因型對於罹患十二指腸潰瘍危險性之比較……….………..…….. 66
表十二(b) 血型 (A型與非A型)與IL-1 B and IL-1 RN基因型對於罹患十二指腸潰瘍危險性之比較……….………..…….. 67
表十三 吸菸習慣與IL-1B和IL-1RN基因型對於罹患十二指腸潰瘍危險性之比較…………………………………………… 68
表十四 飲酒習慣與IL-1B和IL-1RN基因型對可能罹患十二指腸潰瘍危險性之比較
………………………………………………………………… 69
圖目錄
圖一 IL-1基因位置圖……………………………………… 70
圖二 小腸內部構造………………………………………… 71
圖三 小腸內壁……………………………………………… 72
圖四 消化型潰瘍形成途徑…………………………………… 73
圖五 胃部構造…………………………………………… 74
圖六 胃內壁細胞…………………………………………… 75
圖七 胃癌致病過程………………………………………… 76
圖八 進行性胃癌內視鏡分組……………………………… 77
圖九 PCR-CTPP在IL-1B -31位置基因型的電泳圖……… 78
圖十 RFLP在IL-1B -511位置基因型的電泳圖………… 78
圖十一 RFLP在IL-1B +3954位置基因型的電泳圖……… 79
圖十二 PCR在IL-1RN位置基因型的電泳圖…… 79
附錄一 正常人資料………………………………………… 80
附錄二 胃癌病人資料……………………………………… 86
附錄三 十二指腸潰瘍病人資料…………………………… 91
附錄四 haplotype計算方法……………………………… 94
附錄五 Odds Ratio 計算方法…………………………… 97

參考文獻
1. Arend WP, Malyak M, Guthridge CJ et al. Interleukin-1 receptor antagonist: role in biology. Annu Rev Immunol 1998 vol 16: 27-55

2. Beales IL, Calam J. Interleukin 1β and tumour necrosis factor α inhibit acid secretion in cultured rabbit parietal cells by multiple pathways. Gut 1998 vol 42:227-34

3. Beales IL. Effect of Interlukin-1 beta on proliferation of gastric epithelial cells in culture. BMC Gastroenterol 2002 vol 2: 1-8

4. Balkwill FR. Cytokines in cancer therapy. Oxford university press 1989: 150-164

5. Dinarello CA. Biologic Basis for interleukin-1 in disease. Blood 1996 vol 87: 2095-147

6. El-Omar EM, Carrington M, Chow WH et al. Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature 2000 vol 404: 398-402

7. El-Omar E M. The importance of interleukin 1β in Helicobacter pylori associated disease. Gut 2001 vol 48: 743-7

8. Ernst PB, Gold BD. The disease spectrum of Helicobacter pylori: the immunopathogenesis of gastroduodenal ulcer and gastric cancer. Annu Rev Microbiol 2000 vol 54: 615-40
9. Everhart JE. Recent developments in the epidemiology of Helicobacter pylori. Gastroenterol Clin North Am 2000 vol 29: 559-78

10. Garcia-Gonzalez MA, Lanas A, Santolaria S et al. The polymorphism IL-1B and IL-1RN genes in the aetiopathogenesis of peptic ulcer. Clin Exp Immunol 2001 vol 125: 368-75

11. Glickman JN, Antonioli DA. Gastritis. Gastrointest Endosc Clin N Am 2001 vol 11: 717-40

12. Hamajima N, Saito T, Matsuo K et.al. Polymerase chain reaction with confronting two-pair primers for polymorphism genotyping. Jpn J Cancer Res 2000 vol 91: 865-8

13. Hamajima N, Matsuo K, Saito T et al. Interleukin 1 polymorphisms, lifestyle factors, and Helicobacter pylori infection. Jpn J Cancer Res 2001 vol 92: 383-9

14. Hasebe T, Harasawa S, Miwa T. Changes in gastric acid secretion and fasting gastrin levels during healing of duodenal ulcers. Tokai J Exp Clin Med 1995 vol 20: 131-5

15. Lissowska J, Groves FD, Sobin LH et al. Family history and risk of stomach cancer in Warsaw, Poland. Eur J Cancer Prev 1999 vol 8: 223-7

16. Iizuka N, Hazama S, Hirose K et al. Interleukin-1 receptor antagonist mRNA expression and the progression of gastric carcinoma. Cancer Lett 1999 vol 142: 179-84

17. Jain A, Buddhiraja S, Khurana B et al. Risk factors for duodenal ulcer in north India. Trop Gastroenterol 1999 vol 20: 36-9

18. Jung HC, Kim JM, Song IS et al. Helicobacter pylori induced an array of proinflammatory cytokines in human gastric epithelial cell: quantification of mRNA for interleukin-8, -1 alpha/beta, granulocyte-macrophage colony-stimulating factor, monocyte chemoattractant protein-1 and tumour necrosis factor-alpha. J Gastroenterol Hepatol 1997 vol 12: 473-80

19. Kato S, Onda M, Matsukura N et al. Genetic polymorphisms of the cancer related gene and Helicobacter pylori infection in Japanese gastric cancer patients. An age and gender matched case-control study. Cancer 1996 vol 77: 1654-61

20. Kikuchi S. Epidemiology of Helicobacter pylori and gastric cancer. Gastric Cancer 2002 vol 5: 6-15

21. Kodama M, Fujioka T, Murakami K et al. Occurrence of upper gastrointestinal tract disease after Helicobacter pylori eradication. Nippon Rinsho 2001 vol 59: 333-6

22. Landau BR. Essential human antatomy and physiology. Scott, Foresman and Company 1976: 400-421

23. Maier JA, Voulalas P, Roeder D et al. Extensionof the life span of endothelial cells by an interleukin-1α antisense oligomer. Science 1990 vol 249: 1570-4

24. Martini F. Fundamentals of anatomy and physiology. Prentice hall interational inc. 1989: 688-692

25. Miller LC, Isa S, Vannier E, Georgilis K et al. Live Borrelia burgdorferi preferentially active interleukin-1beta gene expression and protein synthesis over the interleukin-1 receptor antagonist. J Clin Invest 1992 vol 90: 906-12

26. Montecucco C, Rappuoli R. Living Dangerously: How Helicobacter pylori Survives in the human stomach. Nat Rev Mol Cell Biol 2001 vol 2: 457-66

27. Nardone G, Staibano S, Rocco A et al. Effect of Helicobacter pylori infection and its eradication on cell proliferation, DNA status, and oncogene expression in patients wih chronic gastritis. Gut 1999 vol 44: 789-99

28. Peek RM Jr, Blaser MJ. Helicobacter Pylori and Gastrointestinal Tract Adenocarcinomas. Nature Rev Cancer 2002 vol 2: 28-37

29. Robert A, Olafsson AS, Lancaster C et al. Interleukin-1 is cytoprotective antisecretory, stimulates PGE2 synthesis by stomach, and retards gastric emptying. Life Sci 1991 vol 48: 123-34

30. Saperas E, Yang H, Tache Y. Interleukin-1β acts at hypothalamic sites to inhibit gastric acid secretion in rats. Am J Physiol 1992 vol 263: G414-8

31. Schulte T, Schols L, Muller T et al. Polymorphisms in interleukin-1 alpha and beta genes and the risk for Parkinson’s disease. Neuroscience Lett 2002 vol 326: 70-72

32. Siewert JR, Klesen D, Maruyama K et al. Gastric cancer diagnosis and treatment. Springer 2000: 2-35

33. Sipponen P, Marshall BJ. Gastritis and gastric cancer. Western countries. Gastroenterol Clin North Am 2000 vol 29: 579-92

34. Staubesand J. Atlas of Human Anatomy, eleventh english edition. Urban and Schwarzenberg 1990 vol 2: 158-182

35. Stevenson FT, Torrano F, Locksley RM et al. Interleukin-1: The patterms of translation and intracellular distribution support alternative secretory mechanisms. J Cell Physiol 1992 vol 152: 223-31

36. Sumitomo M, Tachibana M, Murai M et al. Overexpression of IL-1 ra gene up-reglates interleukin-1β converting enzyme (ICE) gene expression: possible mechanism underlying IL-1β-resistance of cancer cells. Br J Cancer 1999 vol 81: 277-86

37. Tindberg Y, Bengtsson C, Granath F et al. Helicobacter pylori infection in Swedish school children: lack of evidence of child-to-child transmission outside the family. Gastroenterology 2001 vol 121: 310-6
38. Tseng LH, Chen PJ, Lin MT et al. Single nucleotide polymorphisms in intron 2 of the human interleukin-1 receptor antagonist (IL-1Ra) gene: further definition of the IL-1β and IL-1Ra polmorphisms in North American Caucasians and Taiwanese Chinese. Tissue Antigens 2001 vol 57: 318-24

39. Watanabe N, Kobayashi Y. Selective release of a processed form of interleukin-1α. Cytokine 1994 vol 6: 597-601

40. Wilkinson RJ, Patel P, Llewelyn M et al. Influence of polymorphism in the genes for the interleukin (IL)-1 receptor antagonist and IL-1β on tuberculosis. J Exp Med 1999 vol 189: 1863-74

41. You WC, Ma JL, Liu W et al. Blood type and family cancer history in relation to precancerous gastric lesions. Int J Epidemiol 2000 29: 405-7

42. 國家衛生研究院癌症研究組(1999) 台灣癌症臨床研究合作組織出版品：胃癌診治共識。
43. 譚健民翻譯 光復書局編輯部編著(1995) 消化系統，光復書局出版。