跳到主要內容

臺灣博碩士論文加值系統

(44.211.117.197) 您好!臺灣時間:2024/05/21 02:15
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:蔡怡貞
研究生(外文):Yi-Chen Tsai
論文名稱:不同人類膀胱癌細胞株蛋白質的表現
論文名稱(外文):Expression of cellular proteins in different human bladder cancer cell lines
指導教授:邱鐵雄邱鐵雄引用關係
指導教授(外文):Ted H. Chiu
學位類別:碩士
校院名稱:慈濟大學
系所名稱:藥理暨毒理學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:55
中文關鍵詞:膀胱癌
外文關鍵詞:bladder cancer
相關次數:
  • 被引用被引用:2
  • 點閱點閱:660
  • 評分評分:
  • 下載下載:28
  • 收藏至我的研究室書目清單書目收藏:0
膀胱是人體重要的器官之一,在尿液的排泄以及身體恆定上扮演重要的角色。膀胱癌是泌尿系統常見的癌症,而大部分是源自於膀胱移形上皮細胞的病變。早期膀胱癌的症狀跟一般泌尿系統感染很類似,最常見的是無痛性的血尿,因此膀胱癌在早期容易被忽略而難以診斷。此外,膀胱癌的發展通常是多階段性的,所以很難去預測。在本實驗,我們選擇採用四株不同的人類膀胱癌細胞株:BFTC 905、T24、TSGH 8301和RT4,來探討分析膀胱癌可能的生物指標蛋白。實驗利用二維電泳(two-dimensional gel electrophoresis)的方法再配合質譜儀(mass spectrometry)分析可能的指標蛋白質。利用MALDI-TOF 質譜儀鑑定出幾個蛋白質如:prohibitin、eIF5AI、thioredoxin及peroxiredoxin 2 isoform a等當作蛋白質內標。根據這些內標,比較四株細胞株所跑出來的電泳圖,發現有些蛋白質的表現有差異,其中我們已鑑定出17個蛋白質。藉由本實驗不但可以找出膀胱癌可能的生物指標蛋白質,用以作為疾病診斷及預測外,還可建立一個蛋白質鑑定的模式,以分析鑑定細胞中未知的蛋白質。
The urinary bladder plays an important role in urine excretion and homeostasis in human body. Bladder cancer is the common cancer in urinary system. Most of bladder cancer is caused by damage of transitional epithelial cells. Some symptoms of early stage are similar to those of infection; the hematouria with no pain is the common one, so bladder cancer is difficult to be diagnosed at this stage. Besides, bladder cancer development is generally a multistage process. The progression of bladder cancer is difficult to predict. In this study we chose four different human bladder cancer cell lines, BFTC 905, T24, TSGH 8301, and RT4 to determine the possible protein biomarkers for bladder cancer. The expression of cellular proteins was investigated using two-dimensional gel electrophoresis followed by mass spectrometric analysis. Some of the internal standards such as prohibitin, eIF5AI, thioredoxin, and peroxiredoxin 2 isoform a were identified by MALDI-TOF mass spectrometry. Based on internal standards, comparison of the cellular protein profiles of the four cell lines revealed differences in the expression of several proteins. Seventeen proteins were identified in this study. The study may not only reveal the biomarkers of bladder cancer in disease diagnosis and progression prediction, but may also establish an experimental model to identify unknown proteins in those cell lines.
中文摘要.............................................................1 英文摘要.............................................................2 壹、背景介紹..........................................................3 一、癌症.........................................................3 二、膀胱癌........................................................3 1.病理.......................................................4 2.成因.......................................................5 3.症狀.......................................................5 4.診斷.......................................................6 5.治療.......................................................6 6.預後.......................................................7 三、膀胱.........................................................7 1.功能.......................................................7 2.構造.......................................................8 四、蛋白質體學於生物醫學上的應用....................................9 1.基因體學 (genomics)........................................9 2.蛋白質體學(proteomics)......................................9 3.蛋白質體學在癌症醫學上的應用.................................10 貳、研究目的與實驗設計................................................12 一、研究目的.....................................................12 二、實驗設計.....................................................13 參、實驗方法.........................................................14 一、細胞株.......................................................14 二、老鼠體內腫瘤組織..............................................14 三、人類膀胱檢體.................................................14 四、蛋白質的萃取.................................................15 五、蛋白質定量...................................................16 六、二維電泳(Two-dimensional electrophoresis)..................17 七、銀染........................................................18 八、軟體分析.....................................................20 九、膠內酵素分解.................................................20 十、質譜分析.....................................................20 十一、蛋白質鑑定.................................................21 肆、實驗結果.........................................................22 一、膀胱癌細胞株.................................................22 二、老鼠體內腫瘤.................................................23 三、人類膀胱檢體.................................................23 伍、討論............................................................24 陸、結論............................................................35 柒、參考文獻.........................................................36 捌、結果附圖.........................................................43 玖、參考附圖.........................................................54
Abel PD (1988) Prognostic indices in transitional cell carcinoma of the bladder. Br J Urol 62:103-109. Bevec D, Klier H, Holter W, Tschachler E, Valent P, Lottspeich F, Baumruker T, Hauber J (1994) Induced gene expression of the hypusine-containing protein eukaryotic initiation factor 5A in activated human T lymphocytes. Proc Natl Acad Sci U S A 91:10829-10833. Bilim V, Kawasaki T, Katagiri A, Wakatsuki S, Takahashi K, Tomita Y (2000) Altered expression of beta-catenin in renal cell cancer and transitional cell cancer with the absence of beta-catenin gene mutations. Clin Cancer Res 6:460-466. Borer RA, Lehner CF, Eppenberger HM, Nigg EA (1989) Major nucleolar proteins shuttle between nucleus and cytoplasm. Cell 56:379-390. Caraglia M, Tagliaferri P, Budillon A, Abbruzzese A (1999) Post-translational modifications of eukaryotic initiation factor-5A (eIF-5A) as a new target for anti-cancer therapy. Adv Exp Med Biol 472:187-198. Chan WY, Liu QR, Borjigin J, Busch H, Rennert OM, Tease LA, Chan PK (1989) Characterization of the cDNA encoding human nucleophosmin and studies of its role in normal and abnormal growth. Biochemistry 28:1033-1039. Chaves LD, Ostroski BJ, Reed KM (2003) Myosin light chain genes in the turkey (Meleagris gallopavo). Cytogenet Genome Res 102:340-346. Ciocca DR, Oesterreich S, Chamness GC, McGuire WL, Fuqua SA (1993) Biological and clinical implications of heat shock protein 27,000 (Hsp27): a review. J Natl Cancer Inst 85:1558-1570. Cotran RS, Kumar V, Robbins SL (1994) The lower urinary tract. In Robbins Pathologic basis of disease, 5th ed.(Frederick J. Schoen, ed.), W. B. Saunders Company, pp. 991-1005. Dawling S, Roodi N, Mernaugh RL, Wang X, Parl FF (2001) Catechol-O-methyltransferase (COMT)-mediated metabolism of catechol estrogens: comparison of wild-type and variant COMT isoforms. Cancer Res 61:6716-6722. de Graauw M, Tijdens I, Cramer R, Corless S, Timms JF, van de Water B (2005) Heat shock protein 27 is the major differentially phosphorylated protein Involved in renal epithelial cellular stress response and controls focal adhesion organization and apoptosis. J Biol Chem. DeMartino GN, Orth K, McCullough ML, Lee LW, Munn TZ, Moomaw CR, Dawson PA, Slaughter CA (1991) The primary structures of four subunits of the human, high-molecular-weight proteinase, macropain (proteasome), are distinct but homologous. Biochim Biophys Acta 1079:29-38. Di Paolo G, Pellier V, Catsicas M, Antonsson B, Catsicas S, Grenningloh G (1996) The phosphoprotein stathmin is essential for nerve growth factor-stimulated differentiation. J Cell Biol 133:1383-1390. Elledge RM, Green S, Pugh R, Allred DC, Clark GM, Hill J, Ravdin P, Martino S, Osborne CK (2000) Estrogen receptor (ER) and progesterone receptor (PgR), by ligand-binding assay compared with ER, PgR and pS2, by immuno-histochemistry in predicting response to tamoxifen in metastatic breast cancer: a Southwest Oncology Group Study. Int J Cancer 89:111-117. Escobar MA, Hoelz DJ, Sandoval JA, Hickey RJ, Grosfeld JL, Malkas LH (2005) Profiling of nuclear extract proteins from human neuroblastoma cell lines: the search for fingerprints. J Pediatr Surg 40:349-358. Gasdaska JR, Berggren M, Powis G (1995) Cell growth stimulation by the redox protein thioredoxin occurs by a novel helper mechanism. Cell Growth Differ 6:1643-1650. Gavet O, Ozon S, Manceau V, Lawler S, Curmi P, Sobel A (1998) The stathmin phosphoprotein family: intracellular localization and effects on the microtubule network. J Cell Sci 111 (Pt 22):3333-3346. Groblewski GE, Grady T, Mehta N, Lambert H, Logsdon CD, Landry J, Williams JA (1997) Cholecystokinin stimulates heat shock protein 27 phosphorylation in rat pancreas both in vivo and in vitro. Gastroenterology 112:1354-1361. Guo L, Groenendyk J, Papp S, Dabrowska M, Knoblach B, Kay C, Parker JM, Opas M, Michalak M (2003) Identification of an N-domain histidine essential for chaperone function in calreticulin. J Biol Chem 278:50645-50653. Guyton AC, Hall JE (1997) Urine fomation by the kidneys. In Human Physiology and Mechanisms of Disease, 6th (Arthur C. Guyton and John E. Hall eds.), W. B. Saunders Company, pp., 212-235. Hara T, Ishii T, Fujishiro M, Masuda M, Ito T, Nakajima J, Inoue T, Matsuse T (2004) Glutathione S-transferase P1 has protective effects on cell viability against camptothecin. Cancer Lett 203:199-207. Huang CS, Chern HD, Chang KJ, Cheng CW, Hsu SM, Shen CY (1999) Breast cancer risk associated with genotype polymorphism of the estrogen-metabolizing genes CYP17, CYP1A1, and COMT: a multigenic study on cancer susceptibility. Cancer Res 59:4870-4875. Iwaki H, Kageyama S, Isono T, Wakabayashi Y, Okada Y, Yoshimura K, Terai A, Arai Y, Iwamura H, Kawakita M, Yoshiki T (2004) Diagnostic potential in bladder cancer of a panel of tumor markers (calreticulin, gamma -synuclein, and catechol-o-methyltransferase) identified by proteomic analysis. Cancer Sci 95:955-961. Kageyama S, Isono T, Iwaki H, Wakabayashi Y, Okada Y, Kontani K, Yoshimura K, Terai A, Arai Y, Yoshiki T (2004) Identification by proteomic analysis of calreticulin as a marker for bladder cancer and evaluation of the diagnostic accuracy of its detection in urine. Clin Chem 50:857-866. Kanitz MH, Swaminathan S, Savage RE, Jr. (2004) Two-dimensional electrophoretic protein profile analysis following exposure of human uroepithelial cells to occupational bladder carcinogens. Cancer Lett 205:121-131. Kondoh N, Wakatsuki T, Ryo A, Hada A, Aihara T, Horiuchi S, Goseki N, Matsubara O, Takenaka K, Shichita M, Tanaka K, Shuda M, Yamamoto M (1999) Identification and characterization of genes associated with human hepatocellular carcinogenesis. Cancer Res 59:4990-4996. Kristensen P, Johnsen AH, Uerkvitz W, Tanaka K, Hendil KB (1994) Human proteasome subunits from 2-dimensional gels identified by partial sequencing. Biochem Biophys Res Commun 205:1785-1789. Kruse M, Rosorius O, Kratzer F, Bevec D, Kuhnt C, Steinkasserer A, Schuler G, Hauber J (2000) Inhibition of CD83 cell surface expression during dendritic cell maturation by interference with nuclear export of CD83 mRNA. J Exp Med 191:1581-1590. Lebret T, Watson RW, Molinie V, O'Neill A, Gabriel C, Fitzpatrick JM, Botto H (2003) Heat shock proteins HSP27, HSP60, HSP70, and HSP90: expression in bladder carcinoma. Cancer 98:970-977. Lindquist S, Craig EA (1988) The heat-shock proteins. Annu Rev Genet 22:631-677. Liotta LA (1984) Tumor invasion and metastases: role of the basement membrane. Warner-Lambert Parke-Davis Award lecture. Am J Pathol 117:339-348. Liotta LA, Rao CN, Wewer UM (1986) Biochemical interactions of tumor cells with the basement membrane. Annu Rev Biochem 55:1037-1057. Malinoff HL, Wicha MS (1983) Isolation of a cell surface receptor protein for laminin from murine fibrosarcoma cells. J Cell Biol 96:1475-1479. Malinoff HL, McCoy JP, Jr., Varani J, Wicha MS (1984) Metastatic potential of murine fibrosarcoma cells is influenced by cell surface laminin. Int J Cancer 33:651-655. Memon AA, Chang JW, Oh BR, Yoo YJ (2005) Identification of differentially expressed proteins during human urinary bladder cancer progression. Cancer Detect Prev 29:249-255. Nakamura H, Matsuda M, Furuke K, Kitaoka Y, Iwata S, Toda K, Inamoto T, Yamaoka Y, Ozawa K, Yodoi J (1994) Adult T cell leukemia-derived factor/human thioredoxin protects endothelial F-2 cell injury caused by activated neutrophils or hydrogen peroxide. Immunol Lett 42:75-80. Niethammer P, Bastiaens P, Karsenti E (2004) Stathmin-tubulin interaction gradients in motile and mitotic cells. Science 303:1862-1866. Nijtmans LG, de Jong L, Artal Sanz M, Coates PJ, Berden JA, Back JW, Muijsers AO, van der Spek H, Grivell LA (2000) Prohibitins act as a membrane-bound chaperone for the stabilization of mitochondrial proteins. Embo J 19:2444-2451. Nishio K, Nakamura T, Koh Y, Kanzawa F, Tamura T, Saijo N (2001) Oncoprotein 18 overexpression increases the sensitivity to vindesine in the human lung carcinoma cells. Cancer 91:1494-1499. Noh DY, Ahn SJ, Lee RA, Kim SW, Park IA, Chae HZ (2001) Overexpression of peroxiredoxin in human breast cancer. Anticancer Res 21:2085-2090. Park MH, Wolff EC, Folk JE (1993a) Is hypusine essential for eukaryotic cell proliferation? Trends Biochem Sci 18:475-479. Park MH, Wolff EC, Folk JE (1993b) Hypusine: its post-translational formation in eukaryotic initiation factor 5A and its potential role in cellular regulation. Biofactors 4:95-104. Pecina-Slaus N, Pavelic K, Pavelic J (1999) Loss of heterozygosity and protein expression of APC gene in renal cell carcinomas. J Mol Med 77:446-453. Rao NC, Barsky SH, Terranova VP, Liotta LA (1983) Isolation of a tumor cell laminin receptor. Biochem Biophys Res Commun 111:804-808. Reis-Filho JS, Carrilho C, Valenti C, Leitao D, Ribeiro CA, Ribeiro SG, Schmitt FC (2000) Is TTF1 a good immunohistochemical marker to distinguish primary from metastatic lung adenocarcinomas? Pathol Res Pract 196:835-840. Roskams AJ, Friedman V, Wood CM, Walker L, Owens GA, Stewart DA, Altus MS, Danner DB, Liu XT, McClung JK (1993) Cell cycle activity and expression of prohibitin mRNA. J Cell Physiol 157:289-295. Ross DT, Scherf U, Eisen MB, Perou CM, Rees C, Spellman P, Iyer V, Jeffrey SS, Van de Rijn M, Waltham M, Pergamenschikov A, Lee JC, Lashkari D, Shalon D, Myers TG, Weinstein JN, Botstein D, Brown PO (2000) Systematic variation in gene expression patterns in human cancer cell lines. Nat Genet 24:227-235. Sanchez-Font MF, Sebastia J, Sanfeliu C, Cristofol R, Marfany G, Gonzalez-Duarte R (2003) Peroxiredoxin 2 (PRDX2), an antioxidant enzyme, is under-expressed in Down syndrome fetal brains. Cell Mol Life Sci 60:1513-1523. Sasaki M, Yamada Y (1987) The laminin B2 chain has a multidomain structure homologous to the B1 chain. J Biol Chem 262:17111-17117. Schallreuter KU, Pittelkow MR, Wood JM (1986) Free radical reduction by thioredoxin reductase at the surface of normal and vitiliginous human keratinocytes. J Invest Dermatol 87:728-732. Sherr CJ (2004) Principles of tumor suppression. Cell 116:235-246. Silverman DT, Hartge P, Morrison AS, Devesa SS (1992) Epidemiology of bladder cancer. Hematol Oncol Clin North Am 6:1-30. Singh US, Li Q, Cerione R (1998) Identification of the eukaryotic initiation factor 5A as a retinoic acid-stimulated cellular binding partner for tissue transglutaminase II. J Biol Chem 273:1946-1950. Sobel A (1991) Stathmin: a relay phosphoprotein for multiple signal transduction? Trends Biochem Sci 16:301-305. Spector A, Huang RR, Yan GZ, Wang RR (1988) Thioredoxin fragment 31-36 is reduced by dihydrolipoamide and reduces oxidized protein. Biochem Biophys Res Commun 150:156-162. Steglich G, Neupert W, Langer T (1999) Prohibitins regulate membrane protein degradation by the m-AAA protease in mitochondria. Mol Cell Biol 19:3435-3442. Stulik J, Bures J, Jandik P, Langr F, Kovarova H, Macela A (1997) The different expression of proteins recognized by monoclonal anti-heat shock protein 70 (hsp70) antibody in human colonic diseases. Electrophoresis 18:625-628. Timpl R, Rohde H, Robey PG, Rennard SI, Foidart JM, Martin GR (1979) Laminin--a glycoprotein from basement membranes. J Biol Chem 254:9933-9937. Tsui KH, Cheng AJ, Chang PL, Pan TL, Yung BY (2004) Association of nucleophosmin/B23 mRNA expression with clinical outcome in patients with bladder carcinoma. Urology 64:839-844. Varkarakis MJ, Gaeta J, Moore RH, Murphy GP (1974) Superficial bladder tumor. Aspects of clinical progression. Urology 4:414-420. Wang S, Nath N, Adlam M, Chellappan S (1999) Prohibitin, a potential tumor suppressor, interacts with RB and regulates E2F function. Oncogene 18:3501-3510. Wewer UM, Taraboletti G, Sobel ME, Albrechtsen R, Liotta LA (1987) Role of laminin receptor in tumor cell migration. Cancer Res 47:5691-5698. Wimmer K, Kuick R, Thoraval D, Hanash SM (1996) Two-dimensional separations of the genome and proteome of neuroblastoma cells. Electrophoresis 17:1741-1751. Wu MH, Lam CY, Yung BY (1995) Translocation of nucleophosmin from nucleoli to nucleoplasm requires ATP. Biochem J 305 (Pt 3):987-992. Yacoub A, Kelley MR, Deutsch WA (1996) Drosophila ribosomal protein PO contains apurinic/apyrimidinic endonuclease activity. Nucleic Acids Res 24:4298-4303. Yow HK, Wong JM, Chen HS, Lee CG, Davis S, Steele GD, Jr., Chen LB (1988) Increased mRNA expression of a laminin-binding protein in human colon carcinoma: complete sequence of a full-length cDNA encoding the protein. Proc Natl Acad Sci U S A 85:6394-6398. Yun JP, Chew EC, Liew CT, Chan JY, Jin ML, Ding MX, Fai YH, Li HK, Liang XM, Wu QL (2003) Nucleophosmin/B23 is a proliferate shuttle protein associated with nuclear matrix. J Cell Biochem 90:1140-1148. Yung BY, Bor AM, Yang YH (1990) Immunolocalization of phosphoprotein B23 in proliferating and non-proliferating HeLa cells. Int J Cancer 46:272-275. Yung BY, Chang FJ, Bor AM, Lee ES (1992) Schedule-dependent effects of two consecutive, divided, low doses of actinomycin D on translocation of protein B23, inhibition of cell growth and RNA synthesis in HeLa cells. Int J Cancer 52:317-322. Zou Z, Anisowicz A, Hendrix MJ, Thor A, Neveu M, Sheng S, Rafidi K, Seftor E, Sager R (1994) Maspin, a serpin with tumor-suppressing activity in human mammary epithelial cells. Science 263:526-529. 王約翰, 呂福江, 江蓉華, 呂炎田 (1992) 膀胱癌之尿液剝脫細胞學研究及原位移形細胞癌之報告, 膀胱癌治療之最新發展(于大雄, 張聖原, 馬正平)。九州圖書文物有限公司 pp. 3-11. 行政院衛生署 (2004a) http://www.doh.gov.tw/statistic/data/死因摘要/93年/表10.xls. 行政院衛生署 (2004b) http://www.doh.gov.tw/statistic/data/死因摘要/93年/表32.xls. 吳清俊 (1992) 膀胱腫瘤之放射線影像診斷與定期, 膀胱癌治療之最新發展(于大雄, 張聖原, 馬正平)。九州圖書文物有限公司 pp. 13-21. 周綠蘋 (2002) 蛋白質體學於生物醫學上之應用Biological and Biomedical Applications of Proteomics. 生物醫學報導 NTU BioMed. 張正忠 (1978) 生殖泌尿道的腫瘤, 一般泌尿科學(江萬?)。合記圖書出版社 pp. 271-327. 郭漢崇 (2003) 排尿之神經生理學, 臨床尿路動力學。靜思文化志業有限公司 pp. 19-42.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊