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研究生:陳恒立
研究生(外文):Hen-Li Chen
論文名稱:腫瘤壞死因子在戴奧辛致癌過程中可能扮演的角色
論文名稱(外文):Possible role of tumor necrosis factor a in the process of carcinogenesis induced by TCDD
指導教授:王應然王應然引用關係
指導教授(外文):Ying-Jan Wang
學位類別:碩士
校院名稱:國立成功大學
系所名稱:環境醫學研究所
學門:醫藥衛生學門
學類:公共衛生學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:75
中文關鍵詞:氧化性壓力腫瘤壞死因子戴奧辛兩階段動物皮膚促癌實驗
外文關鍵詞:Two Stage Carcinogenesis ExperimentsDioxinTNF-αOxidative Stress State
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戴奧辛(Dioxin)為一系列具有高度毒性及廣泛分布的環境污染物,易經由食入而累積在人體內。在流行病學及實驗動物上皆有證據顯示2,3,7,8-TCDD會導致癌症發生,於1997年被國際癌症組織(International Agency for Research on Cancer, IARC)列為一級的致癌物,但是其致癌機轉仍不清楚。腫瘤壞死因子(Tumor Necrosis Factor-α, TNF-α)是體內調節免疫反應之一個相當重要的因子,目前已知與癌症的發生有關,同時亦有文獻指出TCDD引起的急毒性症狀與TNF-α引起的惡質症相同,且是透過其達到毒性傷害,故本研究欲探討在TCDD慢性致癌過程中TNF-α扮演的角色。
本研究利用兩階段的動物皮膚促癌實驗模式來觀察TCDD的促癌能力及在慢性TCDD暴露的過程中TNF-α在小白鼠體內的變化,以確認其在致癌過程中的重要性,結果發現TCDD無法在小白鼠皮膚促進腫瘤生長,顯示不具有對此品系小鼠皮膚促癌的能力,不過意外地發現小白鼠有肝腫瘤發生;以ELISA測量這些小鼠的血清中TNF-α的濃度發現,小白鼠血清中TNF-α的確受到TCDD誘發,且呈劑量效應關係。另外根據文獻指出,TNF-α的分泌會增加體內氧化性壓力,所以本研究同時進行一個動物灌食實驗模式,觀察灌食TCDD是否會增加體內的氧化性壓力,餵食TCDD後發現隨著TCDD灌食的次數增加,大白鼠體內的氧化性壓力也隨之上升,表示TCDD的暴露會導致氧化性壓力上升。本研究同時建立一個病例對照研究(case-control study),根據TNF-α的基因多型性會影響體內TNF-α分泌的特性(T2 allele會分泌較多的TNF-α),比較肝癌病人與一般族群的基因多型性分佈情形,結果發現TNF-α的分泌量過多與肝癌的危險性增高有相關。最後,我們採用RAW264.7和Hepa1c1兩種細胞進行體外試驗,以RT-PCR分析mRNA的表現,發現只有RAW264.7細胞會被TCDD誘發表現TNF-a。而RAW264.7細胞也會被TCDD誘發釋出自由基。
結合各項實驗的結果,我們推測TCDD引起的慢性致癌過程,是先產生自由基傷害,再藉由誘發肝臟中的巨噬細胞大量表現TNF-a引起大範圍發炎反應,吸引大量免疫細胞浸潤,反覆釋出自由基提高氧化性壓力,最後造成細胞癌化。
Dioxin, a widely distributed environmental contaminant with highly toxicity, can be restored in human body through bioaccumulation. The toxic effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been intensively studied. Epidemiological and experimental studies indicated that exposure of dioxin like chemicals could increase the risk of cancer. Therefore, International Agency for Research on Cancer (IARC) has defined 2,3,7,8-TCDD as a group 1 carcinogen in 1997. However, the precise mechanism of carcinogenic effects of dioxin is unclear. Tumor necrosis factor a (TNF-a) was defined as a key cytokine in inflammatory process, over expression of TNF-a may induce cachexia. It has been reported that TCDD can also induce cachexia. Based on this observation, TNF-a has been postulated as one of the important mediator of TCDD-induced toxicity. In this study, we tried to evaluate the role of TNF-a in the carcinogenic process induced by TCDD.
The mouse skin carcinogenesis model has been used extensively to assess whether chemical and/or physical agents carry a carcinogenic hazard to humans and define the mechanism involved with their carcinogenic effects. We conducted a two-stage carcinogenesis experiment to elucidate the promotional activity of dioxin and found that topical application of TCDD could not induce skin papilloma in CD-1 mice. Surprisingly, we found that dorsal treatment of mice with high dose of TCDD bear liver tumor. Thus, we further examined the serum levels of TNF-a in TCDD-treated mice and found that TCDD could induce TNF-a expression with a dose dependent manner. It has been reported that TCDD could increase oxidative stress in vivo, so we also conducted a rat intubation experiment to evaluate the oxidative stress state of mice exposed to TCDD. The results showed that the oxidative stress of mice increased significantly after continuous intubation of TCDD and decreased mildly to normal level when stopped dosage.
There are three types of gene polymorphism in human TNF-αgene. Different type of inherited TNF-αgene has been proved to possess different gene expressions under stimulation. We conducted a case-control study to determine the association between TNF-αgene polymorphism and human liver cancer. Our results indicated that the proportions of TNF-αgene polymorphism are significantly different in normal population and patients with hepatocellular carcinoma. Finally, as a mechanistic approach, we investigated the possible role of macrophage and hepatic cells in producing TNF-a and oxidative stress in liver tumorigenesis induced by TCDD. The in vitro studies showed that TNF-a gene expression and superoxide releasing could be induced by TCDD in RAW264.7 macrophage cell line but not in Hepa1c1 mice hepatoma cell line. Taken together, TNF-αwas suggested in the present study to play a key role in the liver tumorigenesis induced by TCDD. The releasing of TNF-αand superoxide radical of macrophage result in the elevation of oxidative stress in liver tissue and may finally induces the liver proliferation and tumorigenesis.
中文摘要
Abstract

第一章、序論
第一節、研究動機 1
第二節、研究目的 1

第二章、文獻探討
第一節、戴奧辛簡介 3
第二節、戴奧辛造成的健康效應. 4
第三節、化學致癌的三步驟 5
第四節、腫瘤壞死因子(Tumor Necrosis Factorα, TNF-α)
之生理功能與毒性機制 5
第五節、自由基產生的氧化性傷害與產物 7
第六節、人類腫瘤壞死因子之基因多型性 8

第三章、研究架構
第一節、動物實驗 10
第二節、病例對照研究 12
第三節、細胞實驗 13

第四章、研究材料與方法
第一節、研究材料 14
第二節、兩階段促癌動物實驗 18
第三節、TNF-a (Tumor Necrosis Factor-a) 濃度測定 20
第四節、石蠟切片 (Paraffin section) 21
第五節、蘇木紫伊紅染色 (Hematoxylin & Eosin Staining) 23
第六節、免疫組織染色(Immunohistochemistry )24
第七節、大白鼠灌食實驗 26
第八節、PGF2a (8-epi-prostane f2a)濃度測定 28
第九節、人類TNF-a基因多型性研究 30
第十節、細胞培養 32
第十一節、細胞中TNF-α mRNA的表現 33
第十二節、超氧陰離子自由基(Superoxine anion)測定 35

第五章、研究結果
第一節、兩階段動物皮膚促癌模式實驗 37
第二節、肝臟組織切片染色 39
第三節、TNF-a在小白鼠血清中的表現情形 39
第四節、TCDD大白鼠灌食實驗 40
第五節、人類TNF-a promoter基因多型性研究 40
第六節、TCDD誘發之TNF-a的來源 41
第七節、TCDD誘發氧化性壓力上升情形 42

第六章、討論 43

第七章、參考文獻 49
1.Safe, S.H., Polychlorinated biphenyls (PCBs): environmental impact, biochemical and toxic responses, and implications for risk assessment. Critical Reviews in Toxicology, 1994.24(2) 87-149.2.Van den Berg, M., et al., The toxicokinetics and metabolism of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) and their relevance for toxicity. Critical Reviews in Toxicology, 1994.24(1) 1-74.3.Rappe, D.R.Z.A.C., Environmental sources, distribution, and fate of polychlorinated dibenzodioxins, dibenzofurans, and related organochlorines. In: Dioxins and Health (Schecter A,ed). New York:Plenum Press, 1994.710.4.Schecter, A., Exposure assessment: measurement of dioxins and related chemicals in human tissues. In: Dioxins and Health (Schecter A,ed). New York:Plenum Press, 1994.449-485.5.IARC, IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Vol 69: Polychlorinated Dibenzo-para-Dioxins and Polychlorinated Dibenzofurans. Lyon. International Agency for Research on cancer, 1997.6.Michalek, J.E., et al., Pharmacokinetics of TCDD in veterans of Operation Ranch Hand: 10-year follow-up. Journal of Toxicology & Environmental Health, 1996.47(3) 209-20.7.McConnell, E.E., et al., The comparative toxicity of chlorinated dibenzo-p-dioxins in mice and guinea pigs. Toxicology & Applied Pharmacology, 1978.44(2) 335-56.8.Neal, R.A., et al., The toxicokinetics of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin in mammalian systems. Drug Metabolism Reviews, 1982.13(3) 355-85.9.Kociba, R.J., et al., Results of a two-year chronic toxicity and oncogenicity study of 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats. Toxicology & Applied Pharmacology, 1978.46(2) 279-303.10.Burleson, G.R., et al., Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on influenza virus host resistance in mice. Fundamental & Applied Toxicology, 1996.29(1) 40-7.11.NTP, U.S., Carcinogenesis Bioassay of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (CAS No. 1746-01-6) in OsborneMendel Rats and B6C3F1 Mice (Gavage Study). Tech Rpt No 209. Research Triangle Park, NC:U.S. National Toxicology Program, 1982.12.Abbott, B.D. L.S.B.a.R.M.P., 2,3,7,8- TCDD-induced hyperplasia of the ureteral epithelium produces hydronephrosis in murine fetuses. Teratology, 1987.35(3) 329-34.13.Perdew, G.H., Chemical cross-linking of the cytosolic and nuclear forms of the Ah receptor in hepatoma cell line 1c1c7. Biochemical & Biophysical Research Communications, 1992.182(1) 55-62.14.Pitot, H.C., et al., A method to quantitate the relative initiating and promoting potencies of hepatocarcinogenic agents in their dose-response relationships to altered hepatic foci. Carcinogenesis, 1987.8(10) 1491-9.15.Maronpot, R.R., et al., Dose response for TCDD promotion of hepatocarcinogenesis in rats initiated with DEN: histologic, biochemical, and cell proliferation endpoints. Environmental Health Perspectives, 1993.101(7) 634-42.16.Fingerhut, M.A., et al., Cancer mortality in workers exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin. New England Journal of Medicine, 1991.324(4) 212-8.17.Bertazzi, A., et al., Cancer incidence in a population accidentally exposed to 2,3,7,8-tetrachlorodibenzo-para-dioxin. Epidemiology, 1993.4(5) 398-406.18.Johnson, E.S., Important aspects of the evidence for TCDD carcinogenicity in man. Environmental Health Perspectives, 1993.99 383-90.19.Bishop, C.M. and A.H. Jones, Non-Hodgkin''s lymphoma of the scalp in workers exposed to dioxins. Lancet, 1981.2(8242) 369.20.Klaunig J.E., X.Y., Isenberg J.S., Bachowski S., Kolaja KL., Jiang J., Stevenson DE. and Walborg EF Jr., The role of oxidative stress in chemical carcinogenesis. Environmental Health Perspectives.106 Suppl 1:289-95, 1998.21.Beutler, B.A., The role of tumor necrosis factor in health and disease. Journal of Rheumatology, 1999.26(Suppl 57) 16-21.22.Beutler, B., TNF in pathophysiology: biosynthetic regulation. Journal of Investigative Dermatology, 1990.95(6 Suppl) 81S-84S.23.Beutler, B., I.W. Milsark, and A.C. Cerami, Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science, 1985.229(4716) 869-71.24.Borm, P.J., et al., Spontaneous and stimulated release of tumor necrosis factor-alpha (TNF) from blood monocytes of miners with coal workers'' pneumoconiosis. American Review of Respiratory Disease, 1988.138(6) 1589-94.25.Partanen, R., H. Koskinen, and K. Hemminki, Tumour necrosis factor-alpha (TNF-alpha) in patients who have asbestosis and develop cancer. Occupational & Environmental Medicine, 1995.52(5) 316-9.26.Moore, R.J., et al., Mice deficient in tumor necrosis factor-alpha are resistant to skin carcinogenesis. Nature Medicine, 1999.5(7) 828-31.27.Porat, O., The effect of tumor necrosis factor alpha on the activity of lipoprotein lipase in adipose tissue. Lymphokine Research, 1989.8(4) 459-69.28.Jaattela, M., Biologic activities and mechanisms of action of tumor necrosis factor-alpha/cachectin. Laboratory Investigation, 1991.64(6) 724-42.29.Clark, G.C., et al., Tumor necrosis factor involvement in 2,3,7,8-tetrachlorodibenzo-p-dioxin-mediated endotoxin hypersensitivity in C57BL/6J mice congenic at the Ah locus. Toxicology & Applied Pharmacology, 1991.111(3) 422-31.30.Gorski, J.R., et al., Some endocrine and morphological aspects of the acute toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Toxicologic Pathology, 1988.16(3) 313-20.31.Gasiewicz, T.A. and R.A. Neal, 2,3,7,8-Tetrachlorodibenzo-p-dioxin tissue distribution, excretion, and effects on clinical chemical parameters in guinea pigs. Toxicology & Applied Pharmacology, 1979.51(2) 329-39.32.Brewster, D.W. and F. Matsumura, Differential effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on adipose tissue lipoprotein lipase activity in the guinea pig, rat, hamster, rabbit, and mink. Comparative Biochemistry & Physiology. C, Comparative Pharmacology & Toxicology, 1989.93(1) 49-53.33.Taylor, M.J., et al., Inhibition of acute TCDD toxicity by treatment with anti-tumor necrosis factor antibody or dexamethasone. Toxicology & Applied Pharmacology, 1992.117(1) 126-32.34.Alsharif, N.Z., et al., The effects of anti-TNF-alpha antibody and dexamethasone on TCDD-induced oxidative stress in mice. Pharmacology, 1994.48(2) 127-36.35.Clark, G.C. and M.J. Taylor, Tumor necrosis factor involvement in the toxicity of TCDD: the role of endotoxin in the response. Experimental & Clinical Immunogenetics, 1994.11(2-3) 136-41.36.Suganuma, M., et al., Essential role of tumor necrosis factor alpha (TNF-alpha) in tumor promotion as revealed by TNF-alpha-deficient mice. Cancer Research, 1999.59(18) 4516-8.37.Sies, H., Oxidative stress: from basic research to clinical application. American Journal of Medicine, 1991.91(3C) 31S-38S.38.Delanty, N., et al., 8-Epi PGF2 alpha: specific analysis of an isoeicosanoid as an index of oxidant stress in vivo. British Journal of Clinical Pharmacology, 1996.42(1) 15-9.39.Pitot, H.C., et al., Quantitative evaluation of the promotion by 2,3,7,8-tetrachlorodibenzo-p-dioxin of hepatocarcinogenesis from diethylnitrosamine. Cancer Research, 1980.40(10) 3616-20.40.Stohs, S.J., Oxidative stress induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Free Radical Biology & Medicine, 1990.9(1) 79-90.41.Shertzer, H.G., et al., Dioxin causes a sustained oxidative stress response in the mouse. Biochemical & Biophysical Research Communications, 1998.253(1) 44-8.42.Park, J.Y., M.K. Shigenaga, and B.N. Ames, Induction of cytochrome P4501A1 by 2,3,7,8-tetrachlorodibenzo-p-dioxin or indolo(3,2-b)carbazole is associated with oxidative DNA damage. Proceedings of the National Academy of Sciences of the United States of America, 1996.93(6) 2322-7.43.Ingelman-Sundberg, M. and I. Johansson, Mechanisms of hydroxyl radical formation and ethanol oxidation by ethanol-inducible and other forms of rabbit liver microsomal cytochromes P-450. Journal of Biological Chemistry, 1984.259(10) 6447-58.44.Alsharif, N.Z., T. Lawson, and S.J. Stohs, Oxidative stress induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin is mediated by the aryl hydrocarbon (Ah) receptor complex. Toxicology, 1994.92(1-3) 39-51.45.Abraham, L.J. and K.M. Kroeger, Impact of the -308 TNF promoter polymorphism on the transcriptional regulation of the TNF gene: relevance to disease. Journal of Leukocyte Biology, 1999.66(4) 562-6.46.Kroeger, K.M., K.S. Carville, and L.J. Abraham, The -308 tumor necrosis factor-alpha promoter polymorphism effects transcription. Molecular Immunology, 1997.34(5) 391-9.47.Kroeger, K.M. and L.J. Abraham, Identification of an AP-2 element in the -323 to -285 region of the TNF-alpha gene. Biochemistry & Molecular Biology International, 1996.40(1) 43-51.48.Wilson, A.G., et al., Effects of a tumor necrosis factor (TNF-alpha) promotor base transition on transcriptional activity. British Journal of Rheumatology, 1994.33 89-97.49.Sullivan, K.E., et al., A promoter polymorphism of tumor necrosis factor alpha associated with systemic lupus erythematosus in African-Americans. Arthritis & Rheumatism, 1997.40(12) 2207-11.50.Pociot, F., et al., Association of tumor necrosis factor (TNF) and class II major histocompatibility complex alleles with the secretion of TNF-alpha and TNF-beta by human mononuclear cells: a possible link to insulin-dependent diabetes mellitus. European Journal of Immunology, 1993.23(1) 224-31.51.Messer, G., et al., Allelic variation in the TNF-beta gene does not explain the low TNF-beta response in patients with primary biliary cirrhosis. Scandinavian Journal of Immunology, 1991.34(6) 735-40.52.Cabrera, M., et al., Polymorphism in tumor necrosis factor genes associated with mucocutaneous leishmaniasis. Journal of Experimental Medicine, 1995.182(5) 1259-64.53.Huang, S.L., C.H. Su, and S.C. Chang, Tumor necrosis factor-alpha gene polymorphism in chronic bronchitis. American Journal of Respiratory & Critical Care Medicine, 1997.156(5) 1436-9.54.Chouchane, L., et al., Polymorphism in the tumor necrosis factor-alpha promotor region and in the heat shock protein 70 genes associated with malignant tumors. Cancer, 1997.80(8) 1489-96.55.Hebert, C.D., et al., Relative toxicity and tumor-promoting ability of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,4,7,8-pentachlorodibenzofuran (PCDF), and 1,2,3,4,7,8-hexachlorodibenzofuran (HCDF) in hairless mice. Toxicology & Applied Pharmacology, 1990.102(2) 362-77.56.Berry, D.L., et al., Lack of tumor-promoting ability of certain environmental chemicals in a two-stage mouse skin tumorigenesis assay. Research Communications in Chemical Pathology & Pharmacology, 1978.20(1) 101-8.57.Institute, N.C., DHHS Publication no. NIH80-1757,, 1980.58.Poland, A., D. Palen, and E. Glover, Tumour promotion by TCDD in skin of HRS/J hairless mice. Nature, 1982.300(5889) 271-3.59.Courtney, K.D. and J.A. Moore, Teratology studies with 2,4,5-trichlorophenoxyacetic acid and 2,3,7,8-tetrachlorodibenzo-p-dioxin. Toxicology & Applied Pharmacology, 1971.20(3) 396-403.60.Nagao, T., et al., Tissue distribution after a single subcutaneous administration of 2,3,7,8-tetrabromodibenzo-p-dioxin in comparison with toxicokinetics of 2,3,7,8-tetrachlorodibenzo-p-dioxin in female Wistar rats. Life Sciences, 1996.58(4) 325-36.61.Fan, F., et al., Cytokines (IL-1beta and TNFalpha) in relation to biochemical and immunological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in rats. Toxicology, 1997.116(1-3) 9-16.62.Dietz, D.D., et al., Toxicity studies of acetone administered in the drinking water of rodents. Fundamental & Applied Toxicology, 1991.17(2) 347-60.63.Roberts, L.J., 2nd and J.D. Morrow, The generation and actions of isoprostanes. Biochimica et Biophysica Acta, 1997.1345(2) 121-35.64.Patrono, C. and G.A. FitzGerald, Isoprostanes: potential markers of oxidant stress in atherothrombotic disease. Arteriosclerosis Thrombosis & Vascular Biology, 1997.17(11) 2309-15.65.Connor, M.J., J. Nanthur, and S.M. Puhvel, Influence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on TNF-alpha levels in the skin of congenic haired and hairless mice. Toxicology & Applied Pharmacology, 1994.129(1) 12-5.66.Kock, A., et al., Human keratinocytes are a source for tumor necrosis factor alpha: evidence for synthesis and release upon stimulation with endotoxin or ultraviolet light. Journal of Experimental Medicine, 1990.172(6) 1609-14.67.Gonzalez-Amaro, R., et al., Induction of tumor necrosis factor alpha production by human hepatocytes in chronic viral hepatitis. Journal of Experimental Medicine, 1994.179(3) 841-8.68.Liu, T.Z., et al., Free radical-triggered hepatic injury of experimental obstructive jaundice of rats involves overproduction of proinflammatory cytokines and enhanced activation of nuclear factor kappaB. Annals of Clinical & Laboratory Science, 2001.31(4) 383-90.69.Peters, T.R., J.M. Tosk, and E.A. Goulbourne, Jr., Lucigenin chemiluminescence as a probe for measuring reactive oxygen species production in Escherichia coli. Analytical Biochemistry, 1990.186(2) 316-9.70.Fridovich, I., Superoxide anion radical (O2-.), superoxide dismutases, and related matters. Journal of Biological Chemistry, 1997.272(30) 18515-7.71.Ames, B.N. and L.S. Gold, Too many rodent carcinogens: mitogenesis increases mutagenesis. Science, 1990.249(4972) 970-1.72.Ames, B.N., M.K. Shigenaga, and L.S. Gold, DNA lesions, inducible DNA repair, and cell division: three key factors in mutagenesis and carcinogenesis. Environmental Health Perspectives, 1993.101(Suppl 5) 35-44.73.Weinstein, I.B., Toxicity, cell proliferation, and carcinogenesis. Molecular Carcinogenesis, 1992.5(1) 2-3.74. Berenblum, I., and Shubik, P. A new quantitative approach to the study of the stages of chemical carcinogenesis in the mouse’s skin. Br.J. Cancer 1:383-391,1947
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