請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70291
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 | Hui-Chia Chen | en |
dc.contributor.author | 陳慧珈 | zh_TW |
dc.date.accessioned | 2021-06-17T04:25:21Z | - |
dc.date.available | 2023-09-04 | |
dc.date.copyright | 2018-09-04 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-08-15 | |
dc.identifier.citation | 1. Choo QL, Kuo G, Weiner AJ, Overby LR, Bradley DW, Houghton M. 1989. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359-362.
2. Shimizu YK, Feinstone SM, Kohara M, Purcell RH, Yoshikura H. 1996. Hepatitis C virus: detection of intracellular virus particles by electron microscopy. Hepatology 23:205-209. 3. Robertson B, Myers G, Howard C, Brettin T, Bukh J, Gaschen B, Gojobori T, Maertens G, Mizokami M, Nainan O, Netesov S, Nishioka K, Shin i T, Simmonds P, Smith D, Stuyver L, Weiner A. 1998. Classification, nomenclature, and database development for hepatitis C virus (HCV) and related viruses: proposals for standardization. International Committee on Virus Taxonomy. Arch Virol 143:2493-2503. 4. Moradpour D, Penin F. 2013. Hepatitis C virus proteins: from structure to function. Curr Top Microbiol Immunol 369:113-142. 5. von Schaewen M, Dorner M, Hueging K, Foquet L, Gerges S, Hrebikova G, Heller B, Bitzegeio J, Doerrbecker J, Horwitz JA, Gerold G, Suerbaum S, Rice CM, Meuleman P, Pietschmann T, Ploss A. 2016. Expanding the host range of hepatitis C virus through viral adaptation. Mbio 7. 6. Nouroz F, Shaheen S, Mujtaba G, Noreen S. 2015. An overview on hepatitis C virus genotypes and its control. EJMHG 16:291-298. 7. Kao JH. 2016. Hepatitis C virus infection in Taiwan: Past, present, and future. J Formos Med Assoc 115:65-66. 8. Tellinghuisen TL, Rice CM. 2002. Interaction between hepatitis C virus proteins and host cell factors. Curr Opin Microbiol 5:419-427. 9. Moradpour D, Englert C, Wakita T, Wands JR. 1996. Characterization of cell lines allowing tightly regulated expression of hepatitis C virus core protein. Virology 222:51-63. 10. Drummer HE, Maerz A, Poumbourios P. 2003. Cell surface expression of functional hepatitis C virus E1 and E2 glycoproteins. FEBS Lett 546:385-390. 11. Madan V, Bartenschlager R. 2015. Structural and functional properties of the hepatitis C virus p7 viroporin. Viruses 7:4461-4481. 12. Steinmann E, Penin F, Kallis S, Patel AH, Bartenschlager R, Pietschmann T. 2007. Hepatitis C virus p7 protein is crucial for assembly and release of infectious virions. Plos Pathog 3:962-971. 13. Boukadida C, Fritz M, Blumen B, Fogeron ML, Penin F, Martin A. 2018. NS2 proteases from hepatitis C virus and related hepaciviruses share composite active sites and previously unrecognized intrinsic proteolytic activities. PLoS Pathog 14:e1006863. 14. Pietschmann T, Kaul A, Koutsoudakis G, Shavinskaya A, Kallis S, Steinmann E, Abid K, Negro F, Dreux M, Cosset FL, Bartenschlager R. 2006. Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras. P Natl Acad Sci USA 103:7408-7413. 15. Ashfaq UA, Javed T, Rehman S, Nawaz Z, Riazuddin S. 2011. An overview of HCV molecular biology, replication and immune responses. Virol J 8:161. 16. Kasprzak A, Adamek A. 2008. Role of hepatitis C virus proteins (C, NS3, NS5A) in hepatic oncogenesis. Hepatol Res 38:1-26. 17. Kim JL, Morgenstern KA, Lin C, Fox T, Dwyer MD, Landro JA, Chambers SP, Markland W, Lepre CA, O'Malley ET, Harbeson SL, Rice CM, Murcko MA, Caron PR, Thomson JA. 1996. Crystal structure of the hepatitis C virus NS3 protease domain complexed with a synthetic NS4A cofactor peptide. Cell 87:343-355. 18. Asabe SI, Tanji Y, Satoh S, Kaneko T, Kimura K, Shimotohno K. 1997. The N-terminal region of hepatitis C virus-encoded NS5A is important for NS4A-dependent phosphorylation. J Virol 71:790-796. 19. Kaneko T, Tanji Y, Satoh S, Hijikata M, Asabe S, Kimura K, Shimotohno K. 1994. Production of two phosphoproteins from the NS5A region of the hepatitis C viral genome. Biochem Biophys Res Commun 205:320-326. 20. Neddermann P, Clementi A, De Francesco R. 1999. Hyperphosphorylation of the hepatitis C virus NS5A protein requires an active NS3 protease, NS4A, NS4B, and NS5A encoded on the same polyprotein. J Virol 73:9984-9991. 21. Evans MJ, Rice CM, Goff SP. 2004. Phosphorylation of hepatitis C virus nonstructural protein 5A modulates its protein interactions and viral RNA replication. Proc Natl Acad Sci U S A 101:13038-13043. 22. Tellinghuisen TL, Marcotrigiano J, Gorbalenya AE, Rice CM. 2004. The NS5A protein of hepatitis C virus is a zinc metalloprotein. J Biol Chem 279:48576-48587. 23. Brass V, Bieck E, Montserret R, Wolk B, Hellings JA, Blum HE, Penin F, Moradpour D. 2002. An amino-terminal amphipathic alpha-helix mediates membrane association of the hepatitis C virus nonstructural protein 5A. J Biol Chem 277:8130-8139. 24. Tanji Y, Kaneko T, Satoh S, Shimotohno K. 1995. Phosphorylation of hepatitis C virus-encoded nonstructural protein NS5A. J Virol 69:3980-3986. 25. Gale M, Jr., Blakely CM, Kwieciszewski B, Tan SL, Dossett M, Tang NM, Korth MJ, Polyak SJ, Gretch DR, Katze MG. 1998. Control of PKR protein kinase by hepatitis C virus nonstructural 5A protein: molecular mechanisms of kinase regulation. Mol Cell Biol 18:5208-5218. 26. Tan SL, Nakao H, He Y, Vijaysri S, Neddermann P, Jacobs BL, Mayer BJ, Katze MG. 1999. NS5A, a nonstructural protein of hepatitis C virus, binds growth factor receptor-bound protein 2 adaptor protein in a Src homology 3 domain/ligand-dependent manner and perturbs mitogenic signaling. Proc Natl Acad Sci U S A 96:5533-5538. 27. Zech B, Kurtenbach A, Krieger N, Strand D, Blencke S, Morbitzer M, Salassidis K, Cotten M, Wissing J, Obert S, Bartenschlager R, Herget T, Daub H. 2003. Identification and characterization of amphiphysin II as a novel cellular interaction partner of the hepatitis C virus NS5A protein. J Gen Virol 84:555-560. 28. Macdonald A, Crowder K, Street A, McCormick C, Harris M. 2004. The hepatitis C virus NS5A protein binds to members of the Src family of tyrosine kinases and regulates kinase activity. J Gen Virol 85:721-729. 29. He Y, Staschke KA, Tan SL. 2006. HCV NS5A: A multifunctional regulator of cellular pathways and virus replication. In Tan SL (ed), Hepatitis C Viruses: Genomes and Molecular Biology, Norfolk (UK). 30. He Y, Tan SL, Tareen SU, Vijaysri S, Langland JO, Jacobs BL, Katze MG. 2001. Regulation of mRNA translation and cellular signaling by hepatitis C virus nonstructural protein NS5A. J Virol 75:5090-5098. 31. Wu SC, Chang SC, Wu HY, Liao PJ, Chang MF. 2008. Hepatitis C virus NS5A protein down-regulates the expression of spindle gene Aspm through PKR-p38 signaling pathway. J Biol Chem 283:29396-29404. 32. Lan KH, Sheu ML, Hwang SJ, Yen SH, Chen SY, Wu JC, Wang YJ, Kato N, Omata M, Chang FY, Lee SD. 2002. HCV NS5A interacts with p53 and inhibits p53-mediated apoptosis. Oncogene 21:4801-4811. 33. Behrens SE, Tomei L, De Francesco R. 1996. Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. EMBO J 15:12-22. 34. Cheng JC, Chang MF, Chang SC. 1999. Specific interaction between the hepatitis C virus NS5B RNA polymerase and the 3 ' end of the viral RNA. J Virol 73:7044-7049. 35. Schmidt-Mende J, Bieck E, Hugle T, Penin F, Rice CM, Blum HE, Moradpour D. 2001. Determinants for membrane association of the hepatitis C virus RNA-dependent RNA polymerase. J Biol Chem 276:44052-44063. 36. Pawlotsky JM. 2004. Pathophysiology of hepatitis C virus infection and related liver disease. Trends Microbiol 12:96-102. 37. WHO. 2017. Hepatitis C. http://www.who.int/en/news-room/fact-sheets/detail/hepatitis-c. Accessed 38. Anonymous. 2002. National Institutes of Health Consensus Development Conference Statement: Management of hepatitis C: 2002--June 10-12, 2002. Hepatology 36:S3-20. 39. Liaskou E, Wilson DV, Oo YH. 2012. Innate immune cells in liver inflammation. Mediators Inflamm 2012:949157. 40. Hernandez-Gea V, Friedman SL. 2011. Pathogenesis of liver fibrosis. Annu Rev Pathol 6:425-456. 41. Carulli L, Anzivino C. 2014. Telomere and telomerase in chronic liver disease and hepatocarcinoma. World J Gastroenterol 20:6287-6292. 42. Wiemann SU, Satyanarayana A, Tsahuridu M, Tillmann HL, Zender L, Klempnauer J, Flemming P, Franco S, Blasco MA, Manns MP, Rudolph KL. 2002. Hepatocyte telomere shortening and senescence are general markers of human liver cirrhosis. FASEB J 16:935-942. 43. Satyanarayana A, Manns MP, Rudolph KL. 2004. Telomeres and telomerase: a dual role in hepatocarcinogenesis. Hepatology 40:276-283. 44. Sakamoto M, Effendi K, Masugi Y. 2010. Molecular diagnosis of multistage hepatocarcinogenesis. Jpn J Clin Oncol 40:891-896. 45. Paradis V, Youssef N, Dargere D, Ba N, Bonvoust F, Deschatrette J, Bedossa P. 2001. Replicative senescence in normal liver, chronic hepatitis C, and hepatocellular carcinomas. Hum Pathol 32:327-332. 46. Kawakami Y, Kitamoto M, Nakanishi T, Yasui W, Tahara E, Nakayama J, Ishikawa F, Tahara H, Ide T, Kajiyama G. 2000. Immuno-histochemical detection of human telomerase reverse transcriptase in human liver tissues. Oncogene 19:3888-3893. 47. Hytiroglou P, Kotoula V, Thung SN, Tsokos M, Fiel MI, Papadimitriou CS. 1998. Telomerase activity in precancerous hepatic nodules. Cancer 82:1831-1838. 48. Blackburn EH. 2001. Switching and signaling at the telomere. Cell 106:661-673. 49. Greider CW. 1999. Telomeres do D-loop-T-loop. Cell 97:419-422. 50. Wright WE, Tesmer VM, Huffman KE, Levene SD, Shay JW. 1997. Normal human chromosomes have long G-rich telomeric overhangs at one end. Genes Dev 11:2801-2809. 51. Cong YS, Wright WE, Shay JW. 2002. Human telomerase and its regulation. Microbiol Mol Biol Rev 66:407-425. 52. Feng J, Funk WD, Wang SS, Weinrich SL, Avilion AA, Chiu CP, Adams RR, Chang E, Allsopp RC, Yu J, et al. 1995. The RNA component of human telomerase. Science 269:1236-1241. 53. Takakura M, Kyo S, Kanaya T, Hirano H, Takeda J, Yutsudo M, Inoue M. 1999. Cloning of human telomerase catalytic subunit (hTERT) gene promoter and identification of proximal core promoter sequences essential for transcriptional activation in immortalized and cancer cells. Cancer Res 59:551-557. 54. Kyo S, Takakura M, Taira T, Kanaya T, Itoh H, Yutsudo M, Ariga H, Inoue M. 2000. Sp1 cooperates with c-Myc to activate transcription of the human telomerase reverse transcriptase gene (hTERT). Nucleic Acids Res 28:669-677. 55. Collins K. 2006. The biogenesis and regulation of telomerase holoenzymes. Nat Rev Mol Cell Biol 7:484-494. 56. Gillis AJ, Schuller AP, Skordalakes E. 2008. Structure of the Tribolium castaneum telomerase catalytic subunit TERT. Nature 455:633-637. 57. Nakamura TM, Morin GB, Chapman KB, Weinrich SL, Andrews WH, Lingner J, Harley CB, Cech TR. 1997. Telomerase catalytic subunit homologs from fission yeast and human. Science 277:955-959. 58. Nicholls C, Li H, Wang JQ, Liu JP. 2011. Molecular regulation of telomerase activity in aging. Protein Cell 2:726-738. 59. Li H, Pinto AR, Duan W, Li J, Toh BH, Liu JP. 2005. Telomerase down-regulation does not mediate PC12 pheochromocytoma cell differentiation induced by NGF, but requires MAP kinase signalling. J Neurochem 95:891-901. 60. Allsopp RC, Vaziri H, Patterson C, Goldstein S, Younglai EV, Futcher AB, Greider CW, Harley CB. 1992. Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci U S A 89:10114-10118. 61. Shay JW, Bacchetti S. 1997. A survey of telomerase activity in human cancer. Eur J Cancer 33:787-791. 62. Aravinthan AD, Alexander GJM. 2016. Senescence in chronic liver disease: Is the future in aging? J Hepatol 65:825-834. 63. Ozturk M, Arslan-Ergul A, Bagislar S, Senturk S, Yuzugullu H. 2009. Senescence and immortality in hepatocellular carcinoma. Cancer Lett 286:103-113. 64. Campisi J, d'Adda di Fagagna F. 2007. Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol 8:729-740. 65. Collado M, Medema RH, Garcia-Cao I, Dubuisson ML, Barradas M, Glassford J, Rivas C, Burgering BM, Serrano M, Lam EW. 2000. Inhibition of the phosphoinositide 3-kinase pathway induces a senescence-like arrest mediated by p27Kip1. J Biol Chem 275:21960-21968. 66. Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ. 1993. The P21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75:805-816. 67. Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R, Beach D. 1993. P21 is a universal inhibitor of cyclin kinases. Nature 366:701-704. 68. McConnell BB, Gregory FJ, Stott FJ, Hara E, Peters G. 1999. Induced expression of p16(INK4a) inhibits both CDK4- and CDK2-associated kinase activity by reassortment of cyclin-CDK-inhibitor complexes. Mol Cell Biol 19:1981-1989. 69. Fang L, Igarashi M, Leung J, Sugrue MM, Lee SW, Aaronson SA. 1999. p21Waf1/Cip1/Sdi1 induces permanent growth arrest with markers of replicative senescence in human tumor cells lacking functional p53. Oncogene 18:2789-2797. 70. Beausejour CM, Krtolica A, Galimi F, Narita M, Lowe SW, Yaswen P, Campisi J. 2003. Reversal of human cellular senescence: roles of the p53 and p16 pathways. EMBO J 22:4212-4222. 71. Bringold F, Serrano M. 2000. Tumor suppressors and oncogenes in cellular senescence. Exp Gerontol 35:317-329. 72. Aravinthan A. 2015. Cellular senescence: a hitchhiker's guide. Hum Cell 28:51-64. 73. Parrinello S, Coppe JP, Krtolica A, Campisi J. 2005. Stromal-epithelial interactions in aging and cancer: senescent fibroblasts alter epithelial cell differentiation. J Cell Sci 118:485-496. 74. Plentz RR, Park YN, Lechel A, Kim H, Nellessen F, Langkopf BHE, Wilkens L, Destro A, Fiamengo B, Manns MP, Roncalli M, Rudolph KL. 2007. Telomere shortening and inactivation of cell cycle checkpoints characterize human hepatocarcinogenesis. Hepatology 45:968-976. 75. Huang T-Y. 2009. Molecular mechanisms of HCV NS5A protein involved in the regulation of human telomerase gene. Master thesis. National Taiwan University. 76. Kuo YC, Chen IY, Chang SC, Wu SC, Hung TM, Lee PH, Shimotohno K, Chang MF. 2014. Hepatitis C virus NS5A protein enhances gluconeogenesis through upregulation of Akt-/JNK-PEPCK signalling pathways. Liver Int 34:1358-1368. 77. Nakayama JI, Tahara H, Tahara E, Saito M, Ito K, Nakamura H, Nakanishi T, Tahara E, Ide T, Ishikawa F. 1998. Telomerase activation by hTRT in human normal fibroblasts and hepatocellular carcinomas. Nat Genet 18:65-68. 78. Harley CB, Futcher AB, Greider CW. 1990. Telomeres Shorten during Aging of Human Fibroblasts. Nature 345:458-460. 79. Smirnova OA, Ivanova ON, Bartosch B, Valuev-Elliston VT, Mukhtarov F, Kochetkov SN, Ivanov AV. 2016. Hepatitis C virus NS5A protein triggers oxidative stress by inducing NADPH oxidases 1 and 4 and cytochrome P450 2E1. Oxid Med Cell Longev 2016:8341937. 80. Arima N, Kao CY, Licht T, Padmanabhan R, Sasaguri Y, Padmanabhan R. 2001. Modulation of cell growth by the hepatitis C virus nonstructural protein NS5A. J Biol Chem 276:12675-12684. 81. Agarwal ML, Agarwal A, Taylor WR, Stark GR. 1995. p53 controls both the G2/M and the G1 cell cycle checkpoints and mediates reversible growth arrest in human fibroblasts. Proc Natl Acad Sci U S A 92:8493-8497. 82. Niculescu AB, 3rd, Chen X, Smeets M, Hengst L, Prives C, Reed SI. 1998. Effects of p21(Cip1/Waf1) at both the G1/S and the G2/M cell cycle transitions: pRb is a critical determinant in blocking DNA replication and in preventing endoreduplication. Mol Cell Biol 18:629-643. 83. Ghosh AK, Steele R, Meyer K, Ray R, Ray RB. 1999. Hepatitis C virus NS5A protein modulates cell cycle regulatory genes and promotes cell growth. J Gen Virol 80 ( Pt 5):1179-1183. 84. Qadri I, Iwahashi M, Simon F. 2002. Hepatitis C virus NS5A protein binds TBP and p53, inhibiting their DNA binding and p53 interactions with TBP and ERCC3. Biochim Biophys Acta 1592:193-204. 85. Canepa ET, Scassa ME, Ceruti JM, Marazita MC, Carcagno AL, Sirkin PF, Ogara MF. 2007. INK4 proteins, a family of mammalian CDK inhibitors with novel biological functions. IUBMB Life 59:419-426. 86. Zhang H. 2007. Molecular signaling and genetic pathways of senescence: Its role in tumorigenesis and aging. J Cell Physiol 210:567-574. 87. Herbig U, Jobling WA, Chen BP, Chen DJ, Sedivy JM. 2004. Telomere shortening triggers senescence of human cells through a pathway involving ATM, p53, and p21(CIP1), but not p16(INK4a). Mol Cell 14:501-513. 88. Roberson RS, Kussick SJ, Vallieres E, Chen SY, Wu DY. 2005. Escape from therapy-induced accelerated cellular senescence in p53-null lung cancer cells and in human lung cancers. Cancer Res 65:2795-2803. 89. Yang L, Fang J, Chen J. 2017. Tumor cell senescence response produces aggressive variants. Cell Death Discov 3:17049. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70291 | - |
dc.description.abstract | 目前全球約有七千一百萬人受到C型肝炎病毒 (hepatic C virus;HCV) 的感染,因此HCV感染屬於全球性的健康議題。統計顯示約有85% 急性感染患者會發展成慢性肝病,且10~20% 慢性肝病患者會形成肝硬化,有些患者最終可能惡化成肝細胞癌 (hepatocellular carcinoma;HCC) 而導致死亡。本實驗室先前研究發現肝細胞癌細胞株表達HCV的非結構性蛋白質NS5A時,會造成人類端粒酶逆轉錄酶 (human telomerase reverse transcriptase;hTERT) mRNA表現量下降,故推測端粒酶活性可能也會下降。而已知當細胞的端粒酶活性受到抑制時,細胞最終會走向複製型老化,故本研究想進一步探討NS5A是否會誘導肝細胞癌細胞走向複製型老化。首先檢測HCV的非結構性蛋白質對於hTERT mRNA表現量的影響,由RT-qPCR的結果得知,在表達非結構性蛋白質NS3-5B的HCVR細胞株和表達NS5A的細胞株中hTERT mRNA表現量均會下降,而在表達NS3的細胞株中則不受影響,此結果與先前實驗室的研究成果一致。利用誘導型表達NS5A的不同代數細胞進行細胞老化的相關實驗,由senescence-associated β-galactosidase staining 分析得知表達NS5A的細胞相較於控制組會提前走向複製型老化;且誘導細胞老化的分子p27和p21的表現量上升,其中已知p21參與誘導複製型老化。並檢測NS5A誘導細胞走向複製型老化與hTERT的相關性,結果顯示各代數細胞中的hTERT mRNA表現量下降,且在過度表達hTERT細胞中NS5A誘導的老化細胞數量會減少。此外也在穩定型表達NS5A的細胞確認了NS5A誘導複製型老化的現象。由本研究得知NS5A會抑制hTERT且誘導肝細胞癌細胞走向複製型老化。另外,NS5A會誘導p21的表現。雖已知p21會參與在誘導複製型老化的路徑中,但仍需進一步確認p21在NS5A誘導老化中是否為重要的因子。此外,也得知NS5A會促進p27的表現,推測NS5A所造成的細胞壓力可能會誘導壓力誘導型早期老化,此亦需進一步探討。 | zh_TW |
dc.description.abstract | Globally, an estimated 71 million people have hepatitis C virus (HCV) infection, so it is a health issue in the world. HCV infection causes acute hepatitis, 85% of the cases become chronic, causing chronic hepatitis, cirrhosis (in 10~20% of cases after 10~20 years), and hepatocellular carcinoma (HCC). Previous studies in our laboratory found that HCV non-structural protein 5A (NS5A) caused a decreased hTERT mRNA level in a hepatoma cell line, so it was proposed that telomerase activity may also decrease in NS5A-expressing cells. It is known that a decreased telomerase activity may induce replicative senescence. Therefore, whether NS5A protein could induce cellular senescence by repressing hTERT expression was explored in this study. The effects of NS proteins on hTERT expression were first examined. RT-qPCR data showed that both HCVR cells, which express HCV NS proteins from NS3 to NS5B, and NS5A-expressing cells downregulated hTERT mRNA level. This result is consistent with our previous study. In addition, the phenomenon was not observed in NS3-expressing cells. Next, whether NS5A could induce replicative senescence was examined by using NS5A inducible-expressing cells cultured to different passage numbers. Results from senescence-associated β-galactosidase staining showed that NS5A induced replicative senescence in hepatoma cells earlier than the control group. In addition, the expression of p27 and p21, molecules to induce cellular senescence, were increased. The correlation between NS5A-induced replicative senescence and hTERT was further examined. The data showed that hTERT mRNA level decreased in every passage of the NS5A-expressing cells. The level of senescent cells induced by NS5A was repressed when hTERT was overexpressed, suggesting that NS5A induces cellular senescence by repressing hTERT expression. Replicative senescence was also observed in cells constitutively express NS5A. In conclusion, NS5A repressed hTERT and induced replicative senescence. In addition, NS5A induced p21 expression. Although p21 is known to induce replicative senescence, whether p21 is the predominant mediator in NS5A-induced senescence needs to be further confirmed. Moreover, NS5A also increased p27 expression. Whether NS5A is involved in stress-induced premature senescence needs to be further investigated. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T04:25:21Z (GMT). No. of bitstreams: 1 ntu-107-R05445113-1.pdf: 3987892 bytes, checksum: 79a5b9b64f3878b978da6f5216b2042c (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 國立臺灣大學碩士學位論文口試委員會審定書 i
謝辭 ii 中文摘要 iii 英文摘要 iv 目錄 vi 圖目錄 viii 表目錄 ix 第一章 緒論 1 第一節 C型肝炎病毒 1 第二節 HCV的結構蛋白質 1 第三節 HCV的非結構蛋白質 2 第四節 HCV相關的肝臟疾病與病理機轉 4 第五節 端粒與端粒酶 6 第六節 細胞老化 8 第七節 細胞老化與肝細胞癌症的關係 9 第二章 研究目的 11 第三章 實驗材料 12 第一節 藥品 12 第二節 酵素 13 第三節 抗體 13 第四節 細胞培養液與轉染試劑 14 第五節 套組試劑 14 第六節 其他材料 15 第七節 質體 15 第八節 細胞株 15 第四章 實驗方法 18 第一節 pLenti4-NS5A-V5His的質體構築 (Plasmid construction) 18 第二節 細胞轉型 (Cell transformation) 18 第三節 質體DNA小量製備 (Mini plasmid DNA purification) 19 第四節 DNA瓊脂膠電泳 (Agarose gel electrophoresis) 19 第五節 細胞繼代培養 (Cell subculture) 19 第六節 DNA 轉染 (DNA transfection) 19 第七節 細胞全蛋白質萃取 (Cellular total protein isolation) 20 第八節 蛋白質定量 (Protein quantification) 20 第九節 正十二烷硫酸鈉-聚丙醯胺板膠電泳 (SDS-polyacrylamide gel electrophoresis;SDS-PAGE) 20 第十節 西方墨點法 (Western blotting) 21 第十一節 細胞RNA萃取 (Cellular RNA isolation) 22 第十二節 反轉錄即時定量聚合酶連鎖反應 (Reverse transcription-quantitative polymerase chain reaction;RT-qPCR) 22 第十三節 細胞周期的分析 (Cell cycle analysis) 23 第十四節 老化相關β-半乳糖苷酶染色 (Senescence-associated β-galactosidase staining;SA-β-gal staining) ..23 第十五節 端粒重複序列擴增法 (Telomeric repeat amplification protocol assay;TRAP assay) 23 第五章 實驗結果 25 第一節 表達NS5A的細胞株其hTERT mRNA表現量下降 25 第二節 在Huh7-TR-NS5A細胞株中NS5A的表達會誘導細胞走向複製型老化 25 第三節 NS5A所誘導的複製型老化需依賴hTERT 26 第四節 在穩定型表達NS5A的Huh7細胞株中NS5A會誘導複製型老化 27 第六章 討論 28 圖 31 表 38 參考文獻 39 | |
dc.language.iso | zh-TW | |
dc.title | C型肝炎病毒非結構性蛋白質NS5A參與細胞之複製型老化 | zh_TW |
dc.title | Involvement of hepatitis C virus NS5A protein in replicative senescence | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄧述諄(Shu-Chun Teng),劉旻禕(Min-Yi Liu) | |
dc.subject.keyword | C型肝炎病毒,非結構性蛋白質 5A,人類端粒?逆轉錄,複製型老化, | zh_TW |
dc.subject.keyword | hepatitis C virus (HCV),non-structural protein 5A (NS5A),human telomerase reverse transcriptase (hTERT),replicative senescence, | en |
dc.relation.page | 46 | |
dc.identifier.doi | 10.6342/NTU201803404 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-08-15 | |
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 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-107-1.pdf 目前未授權公開取用 | 3.89 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。