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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張鑫(Shin C. Chang) | |
dc.contributor.author | Shih-Fu Wu | en |
dc.contributor.author | 吳士甫 | zh_TW |
dc.date.accessioned | 2021-06-15T14:08:34Z | - |
dc.date.available | 2020-09-25 | |
dc.date.copyright | 2015-09-25 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-08-19 | |
dc.identifier.citation | Abe, K., Y. Emori, H. Kondo, S. Arai and K. Suzuki (1988). 'The NH2-terminal 21 amino acid residues are not essential for the papain-inhibitory activity of oryzacystatin, a member of the cystatin superfamily. Expression of oryzacystatin cDNA and its truncated fragments in Escherichia coli.' J Biol Chem 263(16): 7655-7659. Appel, N., T. Pietschmann and R. Bartenschlager (2005). 'Mutational analysis of hepatitis C virus nonstructural protein 5A: potential role of differential phosphorylation in RNA replication and identification of a genetically flexible domain.' J Virol 79(5): 3187-3194. Barrett, A. J., H. Fritz, A. Grubb, S. Isemura, M. Jarvinen, N. Katunuma, W. Machleidt, W. Muller-Esterl, M. Sasaki and V. Turk (1986). 'Nomenclature and classification of the proteins homologous with the cysteine-proteinase inhibitor chicken cystatin.' Biochem J 236(1): 312. Barrett, A. J. and H. Kirschke (1981). 'Cathepsin B, cathepsin H, and cathepsin L.' Methods Enzymol 80 Pt C: 535-561. Bjorck, L., A. Grubb and L. Kjellen (1990). 'Cystatin C, a human proteinase inhibitor, blocks replication of herpes simplex virus.' J Virol 64(2): 941-943. Blight, K. J. (2011). 'Charged residues in hepatitis C virus NS4B are critical for multiple NS4B functions in RNA replication.' J Virol 85(16): 8158-8171. Bobek, L. A. and M. J. Levine (1992). 'Cystatins--inhibitors of cysteine proteinases.' Crit Rev Oral Biol Med 3(4): 307-332. Bode, W., R. Engh, D. Musil, U. Thiele, R. Huber, A. Karshikov, J. Brzin, J. Kos and V. Turk (1988). 'The 2.0 A X-ray crystal structure of chicken egg white cystatin and its possible mode of interaction with cysteine proteinases.' EMBO J 7(8): 2593-2599. Bonkovsky, H. L. and S. Mehta (2001). 'Hepatitis C: a review and update.' J Am Acad Dermatol 44(2): 159-182. Bose, S. K., K. Meyer, A. M. Di Bisceglie, R. B. Ray and R. Ray (2012). 'Hepatitis C virus induces epithelial-mesenchymal transition in primary human hepatocytes.' J Virol 86(24): 13621-13628. Chang, S. C., J. C. Cheng, Y. H. Kou, C. H. Kao, C. H. Chiu, H. Y. Wu and M. F. Chang (2000). 'Roles of the AX(4)GKS and arginine-rich motifs of hepatitis C virus RNA helicase in ATP- and viral RNA-binding activity.' J Virol 74(20): 9732-9737. Chang, S. C., J. H. Yen, H. Y. Kang, M. H. Jang and M. F. Chang (1994). 'Nuclear localization signals in the core protein of hepatitis C virus.' Biochem Biophys Res Commun 205(2): 1284-1290. Choo, Q. L., G. Kuo, A. J. Weiner, L. R. Overby, D. W. Bradley and M. Houghton (1989). 'Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome.' Science 244(4902): 359-362. Choo, Q. L., K. H. Richman, J. H. Han, K. Berger, C. Lee, C. Dong, C. Gallegos, D. Coit, R. Medina-Selby, P. J. Barr and et al. (1991). 'Genetic organization and diversity of the hepatitis C virus.' Proc Natl Acad Sci U S A 88(6): 2451-2455. de Chassey, B., V. Navratil, L. Tafforeau, M. S. Hiet, A. Aublin-Gex, S. Agaugue, G. Meiffren, F. Pradezynski, B. F. Faria, T. Chantier and et al (2008). 'Hepatitis C virus infection protein network.' Mol Syst Biol 4: 230. Ding, W. X., H. M. Ni, W. Gao, T. Yoshimori, D. B. Stolz, D. Ron and X. M. Yin (2007). 'Linking of autophagy to ubiquitin-proteasome system is important for the regulation of endoplasmic reticulum stress and cell viability.' Am J Pathol 171(2): 513-524. Erdtmann, L., N. Franck, H. Lerat, J. Le Seyec, D. Gilot, I. Cannie, P. Gripon, U. Hibner and C. Guguen-Guillouzo (2003). 'The hepatitis C virus NS2 protein is an inhibitor of CIDE-B-induced apoptosis.' J Biol Chem 278(20): 18256-18264. Fossum, K. and J. R. Whitaker (1968). 'Ficin and papain inhibitor from chicken egg white.' Arch Biochem Biophys 125(1): 367-375. Friebe, P. and R. Bartenschlager (2002). 'Genetic analysis of sequences in the 3' nontranslated region of hepatitis C virus that are important for RNA replication.' J Virol 76(11): 5326-5338. Gale, M. J., Jr., M. J. Korth, N. M. Tang, S. L. Tan, D. A. Hopkins, T. E. Dever, S. J. Polyak, D. R. Gretch and M. G. Katze (1997). 'Evidence that hepatitis C virus resistance to interferon is mediated through repression of the PKR protein kinase by the nonstructural 5A protein.' Virology 230(2): 217-227. Gogela, N. A., M. V. Lin, J. L. Wisocky and R. T. Chung (2015). 'Enhancing our understanding of current therapies for hepatitis C virus (HCV).' Curr HIV/AIDS Rep 12(1): 68-78. Goffard, A., N. Callens, B. Bartosch, C. Wychowski, F. L. Cosset, C. Montpellier and J. Dubuisson (2005). 'Role of N-linked glycans in the functions of hepatitis C virus envelope glycoproteins.' J Virol 79(13): 8400-8409. Grakoui, A., D. W. McCourt, C. Wychowski, S. M. Feinstone and C. M. Rice (1993). 'Characterization of the hepatitis C virus-encoded serine proteinase: determination of proteinase-dependent polyprotein cleavage sites.' J Virol 67(5): 2832-2843. Grakoui, A., D. W. McCourt, C. Wychowski, S. M. Feinstone and C. M. Rice (1993). 'A second hepatitis C virus-encoded proteinase.' Proc Natl Acad Sci U S A 90(22): 10583-10587. Griffin, S. D., L. P. Beales, D. S. Clarke, O. Worsfold, S. D. Evans, J. Jaeger, M. P. Harris and D. J. Rowlands (2003). 'The p7 protein of hepatitis C virus forms an ion channel that is blocked by the antiviral drug, Amantadine.' FEBS Lett 535(1-3): 34-38. Han, J. H., V. Shyamala, K. H. Richman, M. J. Brauer, B. Irvine, M. S. Urdea, P. Tekamp-Olson, G. Kuo, Q. L. Choo and M. Houghton (1991). 'Characterization of the terminal regions of hepatitis C viral RNA: identification of conserved sequences in the 5' untranslated region and poly(A) tails at the 3' end.' Proc Natl Acad Sci U S A 88(5): 1711-1715. Hawley-Nelson, P., D. R. Roop, C. K. Cheng, T. M. Krieg and S. H. Yuspa (1988). 'Molecular cloning of mouse epidermal cystatin A and detection of regulated expression in differentiation and tumorigenesis.' Mol Carcinog 1(3): 202-211. Hijikata, M., H. Mizushima, T. Akagi, S. Mori, N. Kakiuchi, N. Kato, T. Tanaka, K. Kimura and K. Shimotohno (1993). 'Two distinct proteinase activities required for the processing of a putative nonstructural precursor protein of hepatitis C virus.' J Virol 67(8): 4665-4675. Ito, M., H. Kusunoki and T. Mizuochi (2011). 'Peripheral B cells as reservoirs for persistent HCV infection.' Front Microbiol 2: 177. Jung, M., J. Lee, H. Y. Seo, J. S. Lim and E. K. Kim (2015). 'Cathepsin inhibition-induced lysosomal dysfunction enhances pancreatic beta-cell apoptosis in high glucose.' PLoS One 10(1): e0116972. Kan, Y., T. Okabayashi, S. Yokota, S. Yamamoto, N. Fujii and T. Yamashita (2012). 'Imiquimod suppresses propagation of herpes simplex virus 1 by upregulation of cystatin A via the adenosine receptor A1 pathway.' J Virol 86(19): 10338-10346. Klionsky, D. J., F. C. Abdalla, H. Abeliovich, R. T. Abraham, A. Acevedo-Arozena, K. Adeli, L. Agholme, M. Agnello, P. Agostinis, J. A. Aguirre-Ghiso and et al. (2012). 'Guidelines for the use and interpretation of assays for monitoring autophagy.' Autophagy 8(4): 445-544. Kou, Y. H., M. F. Chang, Y. M. Wang, T. M. Hung and S. C. Chang (2007). 'Differential requirements of NS4A for internal NS3 cleavage and polyprotein processing of hepatitis C virus.' J Virol 81(15): 7999-8008. Kuopio, T., A. Kankaanranta, P. Jalava, P. Kronqvist, T. Kotkansalo, E. Weber and Y. Collan (1998). 'Cysteine proteinase inhibitor cystatin A in breast cancer.' Cancer Res 58(3): 432-436. Lah, T. T., J. L. Clifford, K. M. Helmer, N. A. Day, K. Moin, K. V. Honn, J. D. Crissman and B. F. Sloane (1989). 'Inhibitory properties of low molecular mass cysteine proteinase inhibitors from human sarcoma.' Biochim Biophys Acta 993(1): 63-73. Lerat, H., M. Honda, M. R. Beard, K. Loesch, J. Sun, Y. Yang, M. Okuda, R. Gosert, S. Y. Xiao, S. A. Weinman and S. M. Lemon (2002). 'Steatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus.' Gastroenterology 122(2): 352-365. Liu, Q., R. A. Bhat, A. M. Prince and P. Zhang (1999). 'The hepatitis C virus NS2 protein generated by NS2-3 autocleavage is required for NS5A phosphorylation.' Biochem Biophys Res Commun 254(3): 572-577. Lu, L., Q. Zhang, K. Wu, X. Chen, Y. Zheng, C. Zhu and J. Wu (2015). 'Hepatitis C virus NS3 protein enhances cancer cell invasion by activating matrix metalloproteinase-9 and cyclooxygenase-2 through ERK/p38/NF-kappaB signal cascade.' Cancer Lett 356(2 Pt B): 470-478. Madan, V., D. Paul, V. Lohmann and R. Bartenschlager (2014). 'Inhibition of HCV replication by cyclophilin antagonists is linked to replication fitness and occurs by inhibition of membranous web formation.' Gastroenterology 146(5): 1361-1372 e1361-1369. Moriya, K., H. Fujie, Y. Shintani, H. Yotsuyanagi, T. Tsutsumi, K. Ishibashi, Y. Matsuura, S. Kimura, T. Miyamura and K. Koike (1998). 'The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice.' Nat Med 4(9): 1065-1067. Niederau, C., S. Lange, T. Heintges, A. Erhardt, M. Buschkamp, D. Hurter, M. Nawrocki, L. Kruska, F. Hensel, W. Petry and D. Haussinger (1998). 'Prognosis of chronic hepatitis C: results of a large, prospective cohort study.' Hepatology 28(6): 1687-1695. Pallaoro, M., A. Lahm, G. Biasiol, M. Brunetti, C. Nardella, L. Orsatti, F. Bonelli, S. Orru, F. Narjes and C. Steinkuhler (2001). 'Characterization of the hepatitis C virus NS2/3 processing reaction by using a purified precursor protein.' J Virol 75(20): 9939-9946. Pavlova, A. and I. Bjork (2002). 'The role of the second binding loop of the cysteine protease inhibitor, cystatin A (stefin A), in stabilizing complexes with target proteases is exerted predominantly by Leu73.' Eur J Biochem 269(22): 5649-5658. Pawlotsky, J. M. (2003). 'Mechanisms of antiviral treatment efficacy and failure in chronic hepatitis C.' Antiviral Res 59(1): 1-11. Pileri, P., Y. Uematsu, S. Campagnoli, G. Galli, F. Falugi, R. Petracca, A. J. Weiner, M. Houghton, D. Rosa, G. Grandi and S. Abrignani (1998). 'Binding of hepatitis C virus to CD81.' Science 282(5390): 938-941. Reed, K. E., A. Grakoui and C. M. Rice (1995). 'Hepatitis C virus-encoded NS2-3 protease: cleavage-site mutagenesis and requirements for bimolecular cleavage.' J Virol 69(7): 4127-4136. Reed, K. E. and C. M. Rice (1999). 'Identification of the major phosphorylation site of the hepatitis C virus H strain NS5A protein as serine 2321.' J Biol Chem 274(39): 28011-28018. Ross-Thriepland, D., J. Mankouri and M. Harris (2015). 'Serine phosphorylation of the hepatitis C virus NS5A protein controls the establishment of replication complexes.' J Virol 89(6): 3123-3135. Saitoh, E., S. Isemura, K. Sanada and K. Ohnishi (1991). 'Cystatins of family II are harboring two domains which retain inhibitory activities against the proteinases.' Biochem Biophys Res Commun 175(3): 1070-1075. Sakamuro, D., T. Furukawa and T. Takegami (1995). 'Hepatitis C virus nonstructural protein NS3 transforms NIH 3T3 cells.' J Virol 69(6): 3893-3896. Santolini, E., G. Migliaccio and N. La Monica (1994). 'Biosynthesis and biochemical properties of the hepatitis C virus core protein.' J Virol 68(6): 3631-3641. Sato, T., T. Yamochi, T. Yamochi, U. Aytac, K. Ohnuma, K. S. McKee, C. Morimoto and N. H. Dang (2005). 'CD26 regulates p38 mitogen-activated protein kinase-dependent phosphorylation of integrin beta1, adhesion to extracellular matrix, and tumorigenicity of T-anaplastic large cell lymphoma Karpas 299.' Cancer Res 65(15): 6950-6956. Scarselli, E., H. Ansuini, R. Cerino, R. M. Roccasecca, S. Acali, G. Filocamo, C. Traboni, A. Nicosia, R. Cortese and A. Vitelli (2002). 'The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus.' EMBO J 21(19): 5017-5025. Simmonds, P. (2004). 'Genetic diversity and evolution of hepatitis C virus--15 years on.' J Gen Virol 85(Pt 11): 3173-3188. Sir, D., W. L. Chen, J. Choi, T. Wakita, T. S. Yen and J. H. Ou (2008). 'Induction of incomplete autophagic response by hepatitis C virus via the unfolded protein response.' Hepatology 48(4): 1054-1061. Sir, D., C. F. Kuo, Y. Tian, H. M. Liu, E. J. Huang, J. U. Jung, K. Machida and J. H. Ou (2012). 'Replication of hepatitis C virus RNA on autophagosomal membranes.' J Biol Chem 287(22): 18036-18043. Smith, D. B., J. Bukh, C. Kuiken, A. S. Muerhoff, C. M. Rice, J. T. Stapleton and P. Simmonds (2014). 'Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource.' Hepatology 59(1): 318-327. Stadler, L. K., T. Hoffmann, D. C. Tomlinson, Q. Song, T. Lee, M. Busby, Y. Nyathi, E. Gendra, C. Tiede, K. Flanagan, S. J. Cockell, A. Wipat, C. Harwood, S. D. Wagner, M. A. Knowles, J. J. Davis, N. Keegan and P. K. Ferrigno (2011). 'Structure-function studies of an engineered scaffold protein derived from Stefin A. II: Development and applications of the SQT variant.' Protein Eng Des Sel 24(9): 751-763. Stempniak, M., Z. Hostomska, B. R. Nodes and Z. Hostomsky (1997). 'The NS3 proteinase domain of hepatitis C virus is a zinc-containing enzyme.' J Virol 71(4): 2881-2886. Tai, C. L., W. K. Chi, D. S. Chen and L. H. Hwang (1996). 'The helicase activity associated with hepatitis C virus nonstructural protein 3 (NS3).' J Virol 70(12): 8477-8484. Tedbury, P. R. and M. Harris (2007). 'Characterisation of the role of zinc in the hepatitis C virus NS2/3 auto-cleavage and NS3 protease activities.' J Mol Biol 366(5): 1652-1660. Tong, B., B. Wan, Z. Wei, T. Wang, P. Zhao, Y. Dou, Z. Lv, Y. Xia and Y. Dai (2014). 'Role of cathepsin B in regulating migration and invasion of fibroblast-like synoviocytes into inflamed tissue from patients with rheumatoid arthritis.' Clin Exp Immunol 177(3): 586-597. Tsubouchi, E., S. M. Akbar, N. Horiike and M. Onji (2004). 'Infection and dysfunction of circulating blood dendritic cells and their subsets in chronic hepatitis C virus infection.' J Gastroenterol 39(8): 754-762. Turk, V., J. Brzin, B. Lenarcic, P. Locnikar, T. Popovic, A. Ritonja, J. Babnik, W. Bode and W. Machleidt (1985). 'Structure and function of lysosomal cysteine proteinases and their protein inhibitors.' Prog Clin Biol Res 180: 91-103. Veeravalli, K. K., C. Chetty, S. Ponnala, C. S. Gondi, S. S. Lakka, D. Fassett, J. D. Klopfenstein, D. H. Dinh, M. Gujrati and J. S. Rao (2010). 'MMP-9, uPAR and cathepsin B silencing downregulate integrins in human glioma xenograft cells in vitro and in vivo in nude mice.' PLoS One 5(7): e11583. Wang, L., Y. Tian and J. H. Ou (2015). 'HCV induces the expression of Rubicon and UVRAG to temporally regulate the maturation of autophagosomes and viral replication.' PLoS Pathog 11(3): e1004764. Watashi, K., N. Ishii, M. Hijikata, D. Inoue, T. Murata, Y. Miyanari and K. Shimotohno (2005). 'Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase.' Mol Cell 19(1): 111-122. Welbourn, S. and A. Pause (2007). 'The hepatitis C virus NS2/3 protease.' Curr Issues Mol Biol 9(1): 63-69. Wolk, B., D. Sansonno, H. G. Krausslich, F. Dammacco, C. M. Rice, H. E. Blum and D. Moradpour (2000). 'Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines.' J Virol 74(5): 2293-2304. Yan, S., M. Sameni and B. F. Sloane (1998). 'Cathepsin B and human tumor progression.' Biol Chem 379(2): 113-123. 寇怡衡, 2008. 非結構蛋白質NS3和NS4A參與C型肝炎病毒致病之分子機轉. 國立台灣大學. 蘇堉嘉, 2013. Cystatin-A與C型肝炎病毒非結構性蛋白質NS3之功能性結合作用. 國立臺灣大學. 嚴可芮, 2013. C型肝炎病毒NS3與Integrin 1的交互作用以及對細胞貼附和移動的影響. 國立臺灣大學. 馬瑋健, 2014. Cystatin-A與C型肝炎病毒非結構性蛋白質NS3交互作用之探討. 國立臺灣大學. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/52099 | - |
dc.description.abstract | C型肝炎病毒具有單股正向RNA基因體,可轉譯出一多蛋白質前驅物,並由宿主和病毒蛋白酶切割成結構性與非結構性蛋白質。C型肝炎病毒的蛋白酶有2個,NS2/3半胱胺酸蛋白酶 (cysteine protease)及NS3絲胺酸蛋白酶 (serine protease),兩者共同參與多蛋白質切割,以產生具功能性的非結構性蛋白質,協助病毒RNA的複製。先前研究指出NS3具有致癌性,表現NS3蛋白質可誘發裸鼠產生腫瘤。本實驗室先前的研究中發現基因型1b之病毒NS3蛋白質在輔助因子NS4A存在下具有自我內部截切及細胞轉型能力 ,而且截切產物NS3(1-402)較NS3全長具有更高的細胞轉型能力。當利用親和性管柱分析與NS3(1-402)會形成複合物的細胞因子時,發現Huh7細胞中的cystatin-A可與NS3(1-402)被共同純化出來,表示兩者間可能具有交互作用。而在共同免疫沉澱實驗中也證實了NS3全長與cystatin-A之間確實有交互作用。Cystatin-A為一半胱胺酸蛋白酶抑制蛋白 (cysteine protease inhibitor),可調控細胞中半胱胺酸蛋白酶的活性。當細胞中cystatin-A表現量下降時,可能造成半胱胺酸蛋白酶活性不受調控而導致細胞癌變。本研究在探討NS3與cystatin-A之間交互作用的生物意義及cystatin-A是否參與在NS3造成癌化的過程。結果發現,在Huh7細胞中共同轉染cystatin-A和NS2C-3P蛋白酶表現質體時,並未偵測到NS2/3蛋白酶切割活性有改變,但將細胞中cystatin-A表現抑制時,NS3的表現量會下降。另一方面NS3表現時卻也會使cystatin-A表現量下降,並影響細胞黏著度 (adhesion),而且NS3對細胞黏著度的負調控可藉由外送大量表現的cystatin-A而回復。另外也發現,NS3可以誘發細胞的自噬作用 (autophagy),可能藉此降解cystatin-A。而在利用抑制劑抑制自噬作用後,在cystatin-A表現被抑制的細胞中NS3的表現及自噬作用的活性相較於對照組皆有下降的現象,表示自噬作用可能參與調控NS3蛋白質表現,而其中的機制需更進一步的探討。本研究對於cystatin-A在C型肝炎病毒感染中所扮演的角色提供一個新的研究方向。 | zh_TW |
dc.description.abstract | Hepatitis C virus (HCV) has a positive single-stranded RNA genome. The viral genome can be translated into a polyprotein precursor that is further processed into structure and non-structure proteins by host and virus proteases. HCV possesses two viral proteases, NS2/3 cysteine protease and NS3 serine protease, which participate in the process to generate functional non-structure proteins. Our laboratory has previously demonstrated an internal NS3 cleavage activity of HCV genotype 1b. The major internal cleavage product, NS3(1-402), had a higher transforming activity than the full-length NS3. By performing tandem affinity purification, cystatin-A was co-purified with the NS3(1-402) protein, and its interaction with the full-lenth NS3 was confirmed by co-immunoprecipitation assay. Cystatin-A, as a cysteine protease inhibitor, can modulate cellular cysteine protease activity. The down-regulation of cystatin-A was suggested to be involved in tumorigenesis. In this study, the biological significance for the association between cystatin-A and NS3 and its implication in transformation induced by NS3 were examined. By co-expression NS2C-3P cysteine protease and exogenous cystatin-A in Huh7 cell, no inhibitory effect of cystatin-A to the NS2/3 cysteine protease was observed. On the other hand, NS3 down-regulated expression of cystatin-A protein in various cell types. In addition when cystatin-A as knocked-down, the expression level of NS3 was reduced. Further study demonstrated that NS3 may regulate cystatin-A expression by inducing autophagy. In the presence of inhibitor, NS3 expression was reduced and autophagy activity was significantly diminished in cystatin-A-knockdown cells, suggesting a possible involvement of autophagy in regulation NS3 stability. Furthermore, NS3-mediated reduction on cell adhesion could be rescued by overexpression of cystatin-A, implying a potential role of cystatin-A in HCV pathogenesis. The mechanisms involved in NS3-induced autophagy remain to be elucidated. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T14:08:34Z (GMT). No. of bitstreams: 1 ntu-104-R02445105-1.pdf: 1936309 bytes, checksum: 448afca085c79dfc977ea58420c89c70 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 摘要……………………………………………………………………1 Abstract……………………………………………………………….2 目錄……………………………………………………………………3 緒論……………………………………………………………………6 一、 C型肝炎病毒 二、 C型肝炎病毒的基因體構造及功能 三、 非結構性蛋白質NS2 四、 非結構性蛋白質NS3 五、 半胱胺酸蛋白酶抑制蛋白Cystatin 研究目的………………………………………………………………14 實驗材料………………………………………………………………15 一、 藥品 二、 抗體 三、 細胞培養液及轉染試劑 四、 套組試劑與相關材料 五、 質體 六、 細胞株 實驗方法………………………………………………………………22 一、 瓊脂糖凝膠電泳 (Agarose gel electrophoresis) 二、 細菌轉型 (Transformation) 三、 DNA 轉染 (DNA transfection) 四、 細胞蛋白質之收取 五、 蛋白質定量 六、 十二烷基硫酸鈉-聚丙烯醯胺凝膠電泳 (SDS-polyacrylamide gel electrophoresis, SDS-PAGE) 七、 西方墨點法 (Western blot) 八、 細胞黏著度測試 (Cell adhesion assay) 九、 慢病毒製備與感染 (Lentivirus production and infection) 實驗結果……………………………………………………...……26 一、 Cystatin-A對NS2C-3P(2a)半胱胺酸蛋白酶切割活性的影響 二、 Cystatin-A與NS3表現量的相關性 三、 NS3對cystatin-A蛋白質穩定度的影響 四、 NS3造成cystatin-A表現量下降的作用機制 五、 Cystatin-A可抑制NS3在細胞黏著度的負調控 討論…………………………………………………………………31 一、 Cystatin-A不會抑制NS2C-3P蛋白酶酵素切割的作用 二、 Cystatin-A對NS3蛋白質表現的重要性 三、 Cystatin-A與NS3造成細胞轉型作用的相關性 圖表…………………………………………………………………35 圖一、Cystatin-A對NS2C-3P(2a)半胱胺酸蛋白酶切割活性的影響 圖二、Cystatin-A對NS3蛋白質表現的影響 圖三、NS3在TR-NS3細胞中對cystatin-A蛋白質的影響 圖四、在293T細胞中NS3對內源性cystatin-A蛋白質表現的影響 圖五、在Huh7細胞中NS3對內源性cystatin-A蛋白質表現的影響 圖六、NS3對cystatin-A蛋白質穩定度的影響 圖七、NS3蛋白質在細胞表現時間對cystatin-A蛋白質穩定度的影響 圖八、NS3對cystatin-A蛋白質穩定度的影響與proteosome無關 圖九、NS3與cystatin-A對autophagy的影響 圖十、Cystatin-A回復NS3造成的細胞黏著度的負調控 圖十一、NS3與cystatin-A之交互作用模式圖 圖十二、NS3造成EMT marker的改變 附錄…………………………………………………………………...47 附表一、本研究使用之引子及其序列 附圖一、雞蛋白半胱胺酸蛋白酶抑制蛋白與木瓜蛋白酶交互作用機制圖 參考文獻………………………………………………………………49 | |
dc.language.iso | zh-TW | |
dc.title | C型肝炎病毒非結構性蛋白質NS3對於Cystatin-A蛋白質表現的調控 | zh_TW |
dc.title | Regulation of Cystatin-A Expression by the NS3 Protein of Hepatitis C Virus | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 董馨蓮(Shin-Lian Doong),詹世鵬(Shih-Peng Chan) | |
dc.subject.keyword | C型肝炎病毒,非結構蛋白質NS3,Cystatin-A,自噬作用,細胞黏著度, | zh_TW |
dc.subject.keyword | HCV,NS3,Cystatin-A,Autophagy,Adhesion, | en |
dc.relation.page | 58 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-08-19 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
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