跳到主要內容

臺灣博碩士論文加值系統

(18.97.14.86) 您好!臺灣時間:2025/02/07 22:43
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

我願授權國圖
: 
twitterline
研究生:陳盈利
研究生(外文):Chen Yin-Li
論文名稱:以蛋白質體學的方法鑑定膀胱癌細胞的生物標的分子
論文名稱(外文):To identify biomarkers of bladder cancer by proteomic analysis
指導教授:洪錦堂洪錦堂引用關係
指導教授(外文):Horng Jim-Tong
學位類別:碩士
校院名稱:長庚大學
系所名稱:基礎醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:75
中文關鍵詞:膀胱癌質譜儀細胞膜
外文關鍵詞:bladder cancerDIGEplasma mebrane
相關次數:
  • 被引用被引用:0
  • 點閱點閱:161
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:1
膀胱癌是一種相當普遍的惡性腫瘤,而且其發生率和死亡率有愈來愈高的趨勢,男性發生的機率比女性高出了2.5~5倍。近年來,細胞膜上的膜蛋白質已被廣泛的用為設計藥物之標的物,因此,在病變的細胞中找出大量表達的細胞膜蛋白質將可以提供標的物以設計出治療用抗體或小分子藥物。在這裡,我們欲建立一個簡單且快速的方法,在活細胞的狀況下,直接以螢光標定細胞表面的膜蛋白,再以二維電泳分離膀胱癌的早期細胞株U1及晚期細胞株U4中的膜蛋白質。找到17個在膀胱癌晚期細胞株U1大量表達的點,以質譜儀進行身份鑑定,鑑定出14個蛋白質,其中4個為細胞膜蛋白質,11個為細胞質或粒線體的蛋白質,但是這11個之中有6個是會位移到細胞膜上的蛋白質。從發現可能有差異的蛋白質中,選出確定會位於細胞膜上的蛋白質Annexin II 以及會位移到細胞膜的蛋白質VDAC II進行西方點墨法來做進一步表達量的確認。發現在細胞膜的分離液P2中,U1細胞株內的Annexin II及VDAC II含量皆比U4細胞株中的含量高,但VDAC II並沒有統計上的差異。
Urinary bladder cancer is a common malignancy, and it is also expected to increase in both incidence and mortality. Men are generally 2.5-5 times more likely to develop this malignancy than women. Alterations in the expression of cell surface proteins are often linked to diseases, and surface molecules represent targets for diagnostic and therapeutic approaches. Here we established a new strategy to target cell surface proteins by fluorescence labeling including the possibility of detecting changes in protein abundance between different samples without any prior reduction of the sample complexity. Targeting of cell surface-exposed proteins is labeling of vital cells. It allows detection of fluorophore-tagged cell surface proteins and visualization of the accessible proteome within a single 2-D gel. We analyzed plasma membrane protein expression pattern between two bladder cancer cell lines U1 and U4. U1 is more malignant than U4. We found seventeen spots which were highly expresed in U1 membrane fraction. Fourteen proteins ID were identified by simplifying subsequent MALDI-TOF analysis. Four of these proteins are plasma membrane proteins, and the other ten proteins come from cytosol or mitochondria. According pre-researches, we found six of ten non-plasma membrane proteins would be translocated to plasma membrane. To confirm DIGE results, we chose one plasma membrane protein (Annexin II) and one protein would be translocated to plasma membrane (VDAC II) by werstern blot analysis. We found annexin II and VDAC II were up-regulated in U1 membrane fraction, but the VDAC II has no difference statistically.
目錄
指導教授推薦書………………………………………………………
口試委員會審定書……………………………………………………
授權書…………………………………………………………………iii
誌謝……………………………………………………………………iv
中文摘要………………………………………………………………v
英文摘要………………………………………………………………vi
目錄……………………………………………………………………vii
序論……………………………………………………………………1
材料與方法……………………………………………………………14
結果……………………………………………………………………23
討論……………………………………………………………………31
參考文獻………………………………………………………………38
附表……………………………………………………………………44
附圖……………………………………………………………………51
附錄……………………………………………………………………68
Alban, A., David, S.O., Bjorkesten, L., Andersson, C., Sloge, E., Lewis, S. and Currie, I. (2003) A novel experimental design for comparative two-dimensional gel analysis: two-dimensional difference gel electrophoresis incorporating a pooled internal standard. Proteomics, 3, 36-44.
Alfonso, P., Rivera, J., Hernaez, B., Alonso, C. and Escribano, J.M. (2004) Identification of cellular proteins modified in response to African swine fever virus infection by proteomics. Proteomics, 4, 2037-2046.
Bae, T.J., Kim, M.S., Kim, J.W., Kim, B.W., Choo, H.J., Lee, J.W., Kim, K.B., Lee, C.S., Kim, J.H., Chang, S.Y., Kang, C.Y., Lee, S.W. and Ko, Y.G. (2004) Lipid raft proteome reveals ATP synthase complex in the cell surface. Proteomics, 4, 3536-3548.
Barazi, H.O., Zhou, L., Templeton, N.S., Krutzsch, H.C. and Roberts, D.D. (2002) Identification of heat shock protein 60 as a molecular mediator of alpha 3 beta 1 integrin activation. Cancer Res, 62, 1541-1548.
Bathori, G., Parolini, I., Tombola, F., Szabo, I., Messina, A., Oliva, M., De Pinto, V., Lisanti, M., Sargiacomo, M. and Zoratti, M. (1999) Porin is present in the plasma membrane where it is concentrated in caveolae and caveolae-related domains. J Biol Chem, 274, 29607-29612.
Berggren, K., Chernokalskaya, E., Steinberg, T.H., Kemper, C., Lopez, M.F., Diwu, Z., Haugland, R.P. and Patton, W.F. (2000) Background-free, high sensitivity staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gels using a luminescent ruthenium complex. Electrophoresis, 21, 2509-2521.
Bubenik, J., Baresova, M., Viklicky, V., Jakoubkova, J., Sainerova, H. and Donner, J. (1973) Established cell line of urinary bladder carcinoma (T24) containing tumour-specific antigen. Int J Cancer, 11, 765-773.
Chen, C.J., Chuang, Y.C., You, S.L., Lin, T.M. and Wu, H.Y. (1986) A retrospective study on malignant neoplasms of bladder, lung and liver in blackfoot disease endemic area in Taiwan. Br J Cancer, 53, 399-405.
Chen CJ , K.T., Wu MM . (1988) Arsenic and cancer. The Lancet 20, 414-415.
Chen, K.S., Huang, C.C., Liaw, C.C. and Chen, T.J. (1988) [Multiple primary cancers in blackfoot endemic area: report of a case]. Taiwan Yi Xue Hui Za Zhi, 87, 1125-1128.
Dunn, M.J. (1999) Electroblotting of proteins from 2-D polyacrylamide gels. Methods Mol Biol, 112, 313-318.
FJ Lu, H.G., JS Chiang, CL Hong. (1986) Relationships between the flrorescent intensity of well water and the incidence rate of bladder cancer. . J Chinese Oncol Soc 2, 14-23.
Friedman, D.B., Hill, S., Keller, J.W., Merchant, N.B., Levy, S.E., Coffey, R.J. and Caprioli, R.M. (2004) Proteome analysis of human colon cancer by two-dimensional difference gel electrophoresis and mass spectrometry. Proteomics, 4, 793-811.
Frodin, J.E., Jonsson, E., Moller, T. and Werko, L. (1996) Radiotherapy in Sweden--a study of present use in relation to the literature and an estimate of future trends. Acta Oncol, 35, 967-979.
FS Lin, T.H., TC Tsai, TY Chiu, Jun Chen, TC Hsu, WH Chiang, SW How. (1993) Clinico-patholigical study of bladder tumor. J Urol ROC 4, 1064-1070.
Gade, D., Thiermann, J., Markowsky, D. and Rabus, R. (2003) Evaluation of two-dimensional difference gel electrophoresis for protein profiling. Soluble proteins of the marine bacterium Pirellula sp. strain 1. J Mol Microbiol Biotechnol, 5, 240-251.
Gharbi, S., Gaffney, P., Yang, A., Zvelebil, M.J., Cramer, R., Waterfield, M.D. and Timms, J.F. (2002) Evaluation of two-dimensional differential gel electrophoresis for proteomic expression analysis of a model breast cancer cell system. Mol Cell Proteomics, 1, 91-98.
Golden, J.B. (2003) Prioritizing the human genome: knowledge management for drug discovery. Curr Opin Drug Discov Devel, 6, 310-316.
Goldman, R.C., Trus, B.L. and Leive, L. (1983) Quantitative double-label radiography of two-dimensional protein gels using color negative film and computer analysis. Eur J Biochem, 131, 473-480.
Golijanin, D.J., Kakiashvili, D., Madeb, R.R., Messing, E.M. and Lerner, S.P. (2006) Chemoprevention of bladder cancer. World J Urol, 24, 445-472.
Gorg, A., Obermaier, C., Boguth, G., Harder, A., Scheibe, B., Wildgruber, R. and Weiss, W. (2000) The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis, 21, 1037-1053.
Goshe, M.B., Blonder, J. and Smith, R.D. (2003) Affinity labeling of highly hydrophobic integral membrane proteins for proteome-wide analysis. J Proteome Res, 2, 153-161.
Gygi, S.P., Rist, B., Gerber, S.A., Turecek, F., Gelb, M.H. and Aebersold, R. (1999) Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. Nat Biotechnol, 17, 994-999.
Han, D.K., Eng, J., Zhou, H. and Aebersold, R. (2001) Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry. Nat Biotechnol, 19, 946-951.
Hanash, S. (2003) Disease proteomics. Nature, 422, 226-232.
Hanash, S.M. (2000) Biomedical applications of two-dimensional electrophoresis using immobilized pH gradients: current status. Electrophoresis, 21, 1202-1209.
Hayes, M.J., Shao, D., Bailly, M. and Moss, S.E. (2006) Regulation of actin dynamics by annexin 2. Embo J, 25, 1816-1826.
Hepburn, P.J., and Masters, J. R. W. (1983) The biological characteristics of continuous cell lines derived from human bladder. In: T. Bryan and S. Cohen (eds.), Pathology of Bladder Cancer, 2, 213-227.
Hu, Y., Wang, G., Chen, G.Y., Fu, X. and Yao, S.Q. (2003) Proteome analysis of Saccharomyces cerevisiae under metal stress by two-dimensional differential gel electrophoresis. Electrophoresis, 24, 1458-1470.
Jang, J.H. and Hanash, S. (2003) Profiling of the cell surface proteome. Proteomics, 3, 1947-1954.
Karp, N.A., Kreil, D.P. and Lilley, K.S. (2004) Determining a significant change in protein expression with DeCyder during a pair-wise comparison using two-dimensional difference gel electrophoresis. Proteomics, 4, 1421-1432.
Kernec, F., Unlu, M., Labeikovsky, W., Minden, J.S. and Koretsky, A.P. (2001) Changes in the mitochondrial proteome from mouse hearts deficient in creatine kinase. Physiol Genomics, 6, 117-128.
Kim, B.W., Choo, H.J., Lee, J.W., Kim, J.H. and Ko, Y.G. (2004) Extracellular ATP is generated by ATP synthase complex in adipocyte lipid rafts. Exp Mol Med, 36, 476-485.
Lin, C.W., Lin, J.C. and Prout, G.R., Jr. (1985) Establishment and characterization of four human bladder tumor cell lines and sublines with different degrees of malignancy. Cancer Res, 45, 5070-5079.
Lu, F.J. and Shen, P.M. (1987) [Phthalate esters: chemical carcinogens existing in the water from a well in a blackfoot disease endemic area]. Taiwan Yi Xue Hui Za Zhi, 86, 734-742.
Matsui, N.M., Smith-Beckerman, D.M. and Epstein, L.B. (1999) Staining of preparative 2-D gels. Coomassie blue and imidazole-zinc negative staining. Methods Mol Biol, 112, 307-311.
Mayrhofer, C., Krieger, S., Allmaier, G. and Kerjaschki, D. (2006) DIGE compatible labelling of surface proteins on vital cells in vitro and in vivo. Proteomics, 6, 579-585.
Michener, C.M., Ardekani, A.M., Petricoin, E.F., 3rd, Liotta, L.A. and Kohn, E.C. (2002) Genomics and proteomics: application of novel technology to early detection and prevention of cancer. Cancer Detect Prev, 26, 249-255.
Molloy, M.P. (2000) Two-dimensional electrophoresis of membrane proteins using immobilized pH gradients. Anal Biochem, 280, 1-10.
Petrovich, Z., Baert, L., Boyd, S.D., Brady, L.W., D'Hallewin, M., Heilmann, H.P., Jakse, G., Jones, P.A., Van Der Meijden, A.P., Oyen, R.H., Van Poppel, H., Rotman, M., Sauer, R., Shipley, W.U. and Skinner, E.C. (1998) Management of carcinoma of the bladder. Am J Clin Oncol, 21, 217-222.
Qin, S., Ferdinand, A.S., Richie, J.P., O'Leary, M.P., Mok, S.C. and Liu, B.C. (2005) Chromatofocusing fractionation and two-dimensional difference gel electrophoresis for low abundance serum proteins. Proteomics, 5, 3183-3192.
Rabilloud, T. (1999) Silver staining of 2-D electrophoresis gels. Methods Mol Biol, 112, 297-305.
Rabilloud, T. (2000) Detecting proteins separated by 2-D gel electrophoresis. Anal Chem, 72, 48A-55A.
Rabilloud, T., Gianazza, E., Catto, N. and Righetti, P.G. (1990) Amidosulfobetaines, a family of detergents with improved solubilization properties: application for isoelectric focusing under denaturing conditions. Anal Biochem, 185, 94-102.
Sakai, J., Ishikawa, H., Kojima, S., Satoh, H., Yamamoto, S. and Kanaoka, M. (2003) Proteomic analysis of rat heart in ischemia and ischemia-reperfusion using fluorescence two-dimensional difference gel electrophoresis. Proteomics, 3, 1318-1324.
Seike, M., Kondo, T., Mori, Y., Gemma, A., Kudoh, S., Sakamoto, M., Yamada, T. and Hirohashi, S. (2003) Proteomic analysis of intestinal epithelial cells expressing stabilized beta-catenin. Cancer Res, 63, 4641-4647.
Shaw, J., Rowlinson, R., Nickson, J., Stone, T., Sweet, A., Williams, K. and Tonge, R. (2003) Evaluation of saturation labelling two-dimensional difference gel electrophoresis fluorescent dyes. Proteomics, 3, 1181-1195.
Shevchenko, A., Wilm, M., Vorm, O. and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem, 68, 850-858.
Shin, B.K., Wang, H., Yim, A.M., Le Naour, F., Brichory, F., Jang, J.H., Zhao, R., Puravs, E., Tra, J., Michael, C.W., Misek, D.E. and Hanash, S.M. (2003) Global profiling of the cell surface proteome of cancer cells uncovers an abundance of proteins with chaperone function. J Biol Chem, 278, 7607-7616.
Skynner, H.A., Rosahl, T.W., Knowles, M.R., Salim, K., Reid, L., Cothliff, R., McAllister, G. and Guest, P.C. (2002) Alterations of stress related proteins in genetically altered mice revealed by two-dimensional differential in-gel electrophoresis analysis. Proteomics, 2, 1018-1025.
Spandidos, A. and Rabbitts, T.H. (2002) Sub-proteome differential display: single gel comparison by 2D electrophoresis and mass spectrometry. J Mol Biol, 318, 21-31.
Sugano, K. and Kakizoe, T. (2006) Genetic alterations in bladder cancer and their clinical applications in molecular tumor staging. Nat Clin Pract Urol, 3, 642-652.
Terashima, M., Kim, K.M., Adachi, T., Nielsen, P.J., Reth, M., Kohler, G. and Lamers, M.C. (1994) The IgM antigen receptor of B lymphocytes is associated with prohibitin and a prohibitin-related protein. Embo J, 13, 3782-3792.
Tonge, R., Shaw, J., Middleton, B., Rowlinson, R., Rayner, S., Young, J., Pognan, F., Hawkins, E., Currie, I. and Davison, M. (2001) Validation and development of fluorescence two-dimensional differential gel electrophoresis proteomics technology. Proteomics, 1, 377-396.
Unlu, M., Morgan, M.E. and Minden, J.S. (1997) Difference gel electrophoresis: a single gel method for detecting changes in protein extracts. Electrophoresis, 18, 2071-2077.
Van den Bergh, G., Clerens, S., Cnops, L., Vandesande, F. and Arckens, L. (2003a) Fluorescent two-dimensional difference gel electrophoresis and mass spectrometry identify age-related protein expression differences for the primary visual cortex of kitten and adult cat. J Neurochem, 85, 193-205.
Van den Bergh, G., Clerens, S., Vandesande, F. and Arckens, L. (2003b) Reversed-phase high-performance liquid chromatography prefractionation prior to two-dimensional difference gel electrophoresis and mass spectrometry identifies new differentially expressed proteins between striate cortex of kitten and adult cat. Electrophoresis, 24, 1471-1481.
Verma, M., Wright, G.L., Jr., Hanash, S.M., Gopal-Srivastava, R. and Srivastava, S. (2001) Proteomic approaches within the NCI early detection research network for the discovery and identification of cancer biomarkers. Ann N Y Acad Sci, 945, 103-115.
Von Eggeling, F., Gawriljuk, A., Fiedler, W., Ernst, G., Claussen, U., Klose, J. and Romer, I. (2001) Fluorescent dual colour 2D-protein gel electrophoresis for rapid detection of differences in protein pattern with standard image analysis software. Int J Mol Med, 8, 373-377.
Williams, R.D. (1980) Human urologic cancer cell lines. Invest Urol, 17, 359-363.
Wu, C.C., MacCoss, M.J., Howell, K.E. and Yates, J.R., 3rd. (2003) A method for the comprehensive proteomic analysis of membrane proteins. Nat Biotechnol, 21, 532-538.
Zhou, G., Li, H., DeCamp, D., Chen, S., Shu, H., Gong, Y., Flaig, M., Gillespie, J.W., Hu, N., Taylor, P.R., Emmert-Buck, M.R., Liotta, L.A., Petricoin, E.F., 3rd and Zhao, Y. (2002) 2D differential in-gel electrophoresis for the identification of esophageal scans cell cancer-specific protein markers. Mol Cell Proteomics, 1, 117-124.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top