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研究生:鄒宛玲
研究生(外文):Wan ling Tzou
論文名稱:應用蛋白質體學的技術探討在卵巢癌細胞株中猿病毒腫瘤抗原氮端片段抑制神經膠原致癌基因表現的分子機轉
論文名稱(外文):Proteomics approach to the molecular mechanism of HER2 suppression by SV40 LT425 in ovarian cancer cells
指導教授:高銘欽高銘欽引用關係
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
中文關鍵詞:神經膠原致癌蛋白猿病毒腫瘤抗原氮端區域蛋白質體學
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文獻報告在人類罹患乳癌以及卵巢癌的病患中,約30~40%有神經膠原致癌蛋白 (HER2) 過度表現的情況,且這些病患皆會伴隨著癌細胞轉移以及癒後較差的情況產生。本實驗室過去的研究顯示,在人類卵巢癌細胞株中,猿病毒腫瘤抗原氮端區域 (SV40 LT425) 具有抑制HER2媒介轉形的角色;且從乳癌的細胞株實驗中發現,SV40 LT425不僅會抑制HER2的轉形能力,同時也會抑致癌細胞轉移的能力。然而,SV40 LT425抑制HER2基因表現的機轉仍未清楚,本研究試圖以蛋白質體學技術探討 SV40 LT425 抑制HER2表現的分子機轉。所使用的方法是藉由二維電泳以及基質輔助雷射脫附游離-飛行時間質譜儀去鑑定在SKOV-3.ip1以及 SKOV-3.ip1-LT425-31這兩株細胞株中所存在差異性蛋白質表現的情況。從結果顯示,SV40 LT425的確會造成許多蛋白質的表現量發生變化,例如熱休克蛋白家族、代謝過程中的酵素、Ubiquitination路徑中的調控酵素等。由此類蛋白質的發現,可更深一步地推測SV40 LT425 抑制HER2表現的分子機轉也許是透過影響HER2的降解所導致的。並且發現存在SV40 LT425的卵巢癌細胞會誘發細胞凋亡。
第二章:材料與方法………………………………………………. 13
第一節、實驗材料………………………………………………. 13
壹、細胞株………………………………………………… 13
貳、寡核酸引子(primer)…………………………………… 13
参、化學藥品試劑……………………………………….… 13
肆、主要儀器與器材………………………………………. 15
第二節、實驗方法………………………………………………. 16
壹、 細胞培養與保存…………………………………….
貳、反轉錄-聚合酶連鎖反應 (RT-PCR)…………………. 18
参、DNA電泳……………………………………………. 18
肆、西方點墨法 (Western blotting)……………………… 20
伍、 二維電泳膠法………………………………………. 22
第三章:結果………………………………………………………. 31
第四章:討論………………………………………………………. 35
第五章:結論………………………………………………………. 46
第六章:參考文獻…………………………………………………. 47
表…………………………………………………………………… 56
圖…………………………………………………………………… 62
附錄………………………………………………………………… 80
Aebersold, R. H., Teplow, D. B., Hood, L. E., Kent, S. B. (1986). Electrophoresis onto activated glass:High efficiency preparation of proteins from analytical sodium Dodecyl sulfate polyacrylamide gels for direct sequence analysis. J. Biol. Chem. 261, 4229-4238
Anderson, L., and Anderson, N. G. (1977). High resolution two-dimensional electrophoresis of human plasma proteins. Proc. Natl. Acad. Sci. USA 74, 5421-5425
Bargmann, C. I., Hung, M. C., and Weinberg, R. A. (1986a). The neu oncogene encodes an epidermal growth factor receptor-related protein. Nature 319, 226-230
Bhattacharya, R., and Cabral, F. (2004). A ubiquitous β-tubulin disrupts microtubule assembly and inhibits cell proliferation. Molecular Biology of the Cell 15,2123-1231
Blackstock, W. P., and Weir, M. P. (1999). Proteomics:quantitative and physical mapping of cellular proteins. Trends Biotechnol. 17, 121-127
Brown, M., McComack, M., Zinn, K. G., Farrell, M. P., Bikel, I., and Livingston, D. M. (1986). A recombinant murine retrovirus for simian virus 40 large T cDNA transforms mouse fibroblasts to anchorage-independent growth. J Virol 60, 290-293
Burnette, W. N. (1981). Western blotting:Electrophoretic transfer of proteins form sodium dodecyl sulfate-polyacrylamide gels tounmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal. Biochem. 112, 195-203
Cande, C., Vahsen N., Kouranti I., Schmitt E., Daugas E., Spahr C., Luban J., Kroemer, R. T., Giordanetto, F., Garrido, C., Penninger J. M., and Kroemer G. (2004). AIF and cyclophilin A cooperate in apoptosis-associated chromatinolysis. Oncogene 23, 1514–1521
Carraway, K. L., and Cantley, L. C. (1994). A neu acquaintance for erbB3 and erbB4:a role for receptor heterodimerization in growth signaling. Cell 78, 5-8
Challberg, M. D., and Kelly, T. J. (1989). Animal virus DNA replication. Annu Rev Biochem 58, 671-717
Chuang, T. C., Lee, Y. J., Liu, J. Y., Lin, Y. S., Li, J. W., Wang, V., Law, S. L., and Kao, M. C. (2003). EBNA1 may prolong G2/M phase and sensitize HER2/neu-overexpressing ovarian cancer cells to both topoisomerase II-targeting and paclitaxel drug. Biochem Biophys Res Commun 307, 653-659
Chuang, T. C., Yu, Y. H., Lin, Y. S., Wang, S. S., and Kao, M. C. (2002). The N-terminal domain of SV40 large T antigen represses the HER2/neu-mediated transformation and metastatic potential in breast cancers. FEBS Lett 511, 46-50
Chua1, B.T., Volbracht C., Tan, K. O., Li, R., Yu, V. C., and Li. Peng. (2003). Mitochondrial translocation of cofilin is an early step in apoptosis induction. NATURE CELL BIOLOGY 5,1083-1093
Cleveland, D.W., Fischer, S.G., Kirschner, M.W. & Laemmli, U.K. (1977). Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J. Biol. Chem. 252, 1102-1106
Coussens, L., Yang-Feng, T. L., Liao, Y. C., Chen, E., Gray, A., McGrath, J., Seeburg, P. H., Libermann, T. A., Schlessinger, J., Francke, U., and et al. (1985). Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science, 230, 1132-1139
Dick, D. M., and Richard, I. M. (2004). Molecular chaperones and the stress of oncogenesis. Oncogene 23, 2907–2918
Fanning, E., and Knippers, R. (1992). Structure and function of simian virus 40 large tumor antigen. Annu Rev Biochem 61, 55-85
Fiers, W., Contreras, R., Haegemann, G., Rogiers, R., Van de Voorde, A., Van Heuverswyn, H., Van Herrweghe, J., Volckaert, G., and Ysebaert, M. (1978). Complete nucleotide sequence of SV40 DNA. Nature 273, 113-120
Fujita, J., Bandoh1, S., Yang1Y., Wu1, F., Ohtsuki, Y., Yoshinouchi, T., and Ishida, T. (2003). High Molecular Weight Vimentin Complex Is Formed after Proteolytic Digestion of Vimentin by Caspase-3: Detection by Sera of Patients with Interstitial Pneumonia. Microbiol. Immunol., 47, 447–451
Fukushige, S., Matsubara, K., Yoshida, M., Suzuki, T., Semba, K., Toyoshima, K., and Yamamoto, T. (1986). Localization of a novel v-erbB-related gene, c-erbB-2, on human chromosome 17 and its amplification in a gastric cancer cell line. Mol Cell Biol 6, 955-958
Galit, E. S., Haklai, R., Ballan, E., Gabius H. J., and Kloog Y. (2002). Galectin-1 Augments Ras Activation and Diverts Ras Signals to Raf-1 at the Expense of Phosphoinositide 3-Kinase. Journal of biological chemistry 277, 37169–37175
Gonnet, F., Lemaître, G., Waksman, G. and Tortajada, J. (2003). MALDI/MS peptide mass fingerprinting for proteome analysis: identification of hydrophobic proteins attached to eucaryote keratinocyte cytoplasmic membrane using different matrices in concert. Proteome Science 1,1-7
Graus-Porta, D., Beerli, R. R., Daly, J. M., and Hynes, N. E. (1997). ErbB-2, the preferred heterodimerization partner of all ErbB receptors, is a mediator of lateral signaling. Embo J 16, 1647-1655
Gulewitsch W., and Amiradzibi S. (1900). Ber. Deut. Chem. Ges. 33, 1902-1903
Helmbrecht, K., Zeise, E., and Rensing, L. (2000). Chaperones in cell cycle regulation and mitogenic signal transduction: a review. Cell Prolif. 33, 341-365
Henriette J. G. Arts , Harry H. , Willy L. , Pax H. B. Willemse , Elisabeth G. E. De Vries , Harm H. Kampinga , Ate G.J. Van der Zee (1999). HSP27 expression in ovarian carcinoma: Relation in response to chemotherapy and prognosis. Int J Cancer 84, 234-238
Henzel, W. J., Billeci, T. M., Stults, J. T., Wong, S. C., Grimley, C. & Watanabe, C. (1993). Identifying proteins from two-dimensional gels by molecular mass searching of peptide fragments in protein sequence databases. Proc. Natl. Acad. Sci. USA 90, 5011-5015
Hong, S. J., Dawson,T. M., and Dawson, V. L. (2004). Nuclear and mitochondrial conversations in cell death: PARP-1 and AIF signaling TRENDS in Pharmacological Sciences 25, 259-264
Hung, M. C., Matin,A., Zhang, Y., Xing, X., Sorgi, F., Huang, L., and Yu, D. (1995). HER-2/neu-targeting gene therapy-a review. Gene 159,65-71
Janke J., Schlüter K., Burkhard Jandrig, Theile, M., Kölble, K., Arnold, W., Grinstein, E., Schwartz, A., Lope, E. S., Schlag, P. M., Jockusch B. M., and Scherneck, S.(2000). Suppression of Tumorigenicity in Breast Cancer Cells by the Microfilament Protein Profilin 1. J. Exp. Med. 191, 1675–1685
Jorg, H. W. Distler, Wenger, R. H., Gassmann, M., Kurowska, M., Hirth, A., Gay, S.,1 and Distler, O. (2004). Physiologic Responses to Hypoxia and Implications for Hypoxia-Inducible Factors in the Pathogenesis of Rheumatoid Arthritis. ARTHRITIS & RHEUMATISM 50, 10-23
Kaltschmidt E., and Wittmann H. G. (1970). Ribosomal protein VII, Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal. Biochem. 36, 401-412
Kao, M. C., Liu, G. Y., Chuang, T. C., Lin, Y. S., Wuu, J. A., and Law, S. L. (1998a). The N-terminal 178 amino acid domain only the SV40 large T antigen acts as a transforming suppressor of the HER-2/neu oncogene. Oncogene 16,547-554
Klose, J., Nock, C., Herrmann M., Stuhler K., Marcus K., Bluggel M., Krause E., Schalkwyk L. C., Rastan S., Brown S. D., Bussow K. Himmelbauer H., and Lehrach H. (2002). Genetic analysis of the mouse brain proteome. Nat Genet 30, 385-393
Kriegler, M., Perez, C. F., Hardy, C., and Botchan, M. (1984). Transformation mediated by the SV40 T antigens:separation of the overlapping SV40 early genes with a retroviral vector. Cell 38, 483-491
Lin, Y. C., Peng, J. M., and Wang, W. B. (2000). The N-terminal common domain of simian virus 40 large T and small T antigens acts as a transformation suppressor of the HER-2/neu oncogene. Oncogene 19, 2704-2713
Manfredi, J. J., Prives, C. (1994). The transforming activity of simian virus 40 large tumor antigen. Biochem Biophys Acta 1198, 65-83
Matin, A., and Hung, M. C. (1993). Negative regulation of the neu promoter by the SV40 large T antigen. Cell Growth Differ 4, 1051-1056
Matin, A., and Hung, M. C. (1994). The retinoblastoma gene product, Rb, represses neu expression through two regions within the neu regulatory sequence. Oncogene 9, 1333-1339
Matsudaira P. (1987). Sequence form picomole quantities of proteins electroblotting onto polyvinylidene difluoride membranes. J. Biol. Chem. 262, 10035-10038
O’Farrell P. H. (1975). High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250,4007-4021
Padhy, L. C., Shih, C., Cowing, D., Finkelstein, R., and Weinberg, R. A. (1982). Identification of a phosphorylation specially induced by the transforming DNA of rat neuroblastomas. Cell 28, 865-871
Pandey, A., and Mann M. (2000). Proteomics to study genes and genomics. Nature 405, 837-846
Petit, C. A., Gardes, M., and Feunteun, J. (1983). Immortalization of rodent embryo fibroblasts by SV40 is maintained by the A gene. Virology 127, 74-82
Plowman, G. D., Green, J. M., Culocuscou, J. M., Carlton, G. W., Rothwell, V. M., and Buckley, S. (1993). Heregulin induces tyrosine phosphorylation of HER4/p180 erbB4. Nature 366, 473-475
Reddy, V. B., Thimmappaya, B., Dhar, R., Subramanian, K. N., Zain, B.
S., Pan, J., Ghosh, P. K., Celma, M. L., and Weissman, S. M. (1978).
The genome of simian virus 40. Science 200, 494-502
Reimertz, C., Kögel, D., Rami, A, Chittenden Thomas,and Jochen H.M. Prehn.(2003). Gene expression during ER stress–induced apoptosis in neurons: induction of the BH3-only protein Bbc3/PUMA and activation of the mitochondrial apoptosis pathway.The Journal of Cell Biology 162, 587–597
Sanger F., and Tuppy H. (1953). The amino acid sequence in the phenylalanyl chain of insulin. Biochem. J. 49, 463-490
Schechter, A. L., Hung, M. C., Vaidyanathan, L., Weinberg, R. A., Yang-Feng, T. L., Francke, U., Ullrich, A., and Coussens, L. (1985). The neu gene:an erbB-homologous gene distinct from and unliked to the gene encoding the EGF receptor. Science 229, 976-978
Shapiro A. L., Vinuela E., and Maizel JV Jr. (1967). Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biophys Res Commun 28, 815-820
Shih, C., Padhy, L. C., Murray, M., and Weinberg, R. A. (1981). Transforming genes of carcinomas and neuroblastomas introduced into mouse fibroblasts. Nature 290, 261-264
Slamon, D. J., Clark, G M., Wong, S. G., Levin, W. J., Ulrich, A., and McGuire, W. L. (1987). Human breast cancer:correlation of relapse and survival with amplication of the HER-2/neu oncogene. Science 235, 177-182
Spengler B., and Kaufmann R. (1993). Mass Spectrometric sequencing of linear peptides by product-ion analysis in a reflectron time of flight mass spectrometer using matrix-assisted laser desorption ionization. Rapid Commum Mass Spectrom 7, 902-910
Stern, D. F., Heffernan, P. A., and Weinberg, R. A. (1986). p185, a product of the neu proto-oncogene, is a receptor-like protein associated with tyrosine kinase activity. Mol Cell Biol 6, 1729-1740
Steven, X., Sophie, R., Dita, R., Trevor, H., Yves, A., Robin, C., Paul, T., Rejean, R., Antony, R., and Donald, W. N. (1999). Hsp60 accelerates the maturation of pro-caspase-3 by upstream activator proteases during apoptosis. EMBO Journal 18,2049-2056
Suen, T. C., and Hung, M. C. (1991). c-myc reverses neu-induced transformed morphology by transcriptional repression. Mol Cell Biol 11, 354-362
Tan, M., Yao, J., and Yu, D. (1997). Overexpression of the c-erbB-2 gene enhanced intrinsic metastasis potential in human breast cancer cells without increasing their transformation abilities. Cancer Res 57, 1199-1205
Towbin, H., Staehelin, T., and Gordon, J. (1979). Electrophoretic transfer of proteins form polyacrylamide gels to nitrocellulose sheet:procedure and some application. Proc. Natl. Acad. Sci. USA 76, 4350-4354
Vandekerckhove, J., Bauw, G., Puype, M., Van, D. J., and Van, M. M. (1985). Protein-blotting on polybrene-coated glass-fiber sheets. A basis of acid hydrolysis and gas-phase sequencing of picomole quantities of protein previously separated on SDS-polyacrylamide gel. Eur. J. Biochem. 152, 9-19
Xu, W., Edward, G. M., Kim, J. S., Trepel, J. B., and Neckers, L. M. (2002). Hsp90, not Grp94, regulates the intracellular trafficking and stability of nascent ErbB2. Cell Stress & Chaperones. 7, 91–96
Wittmann-Liebold B., and Wittmann H. G. (1964). Die primare Proteinstruktur von Stammen des Tabakmosaikvirus Aminosauresequenzen des Proteins des TMV-Stamme’s dahlemense. (1964). Teil I. Hoppe-Seylers Z. Physiol. Chem. 335, 69-116
Wittmann H. G., and Wittmann-Liebold B. (1974). Chemical structure of bacterical ribosomal proteins. Cold Spring Harbor Symp Quant. Biol 115-140
Yamamoto, T., Ikawa, S., Akiyama, T., Semba, K., Nomura, N., Miyajima, N., Saito, T., and Toyoshima, K. (1986). Similarity of protein encoded by the human c-erb-B2 gene to epidermal growth factor receptor. Nature 319, 230-234
Yu, D., and Hung, M. C. (2000). Overexpression of ErbB2 in cancer and ErbB2-targeting strategies. Oncogene 19, 6115-6121
Yu, D., Liu, B., Tan, M., Li, J., Wang, S. S., and Hung, M. C. (1996). Overexpression of c-erbB-2/neu in breast cancer cells confers increased resistance to Taxol via mdr-1-independent mechanisms. Oncogene 13, 1359-1365
Yu, D., Wolf, J. K., Scanlon, M., Price, J. E., and Hung, M. C. (1993). Enhanced c-erbB-2/neu expression in human ovarian cancer cells correlates with more severe malignancy that can be suppressed by E1A. Cancer Res 53, 891-898
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