1887

Abstract

Chronic hepatitis C virus (HCV) infection results in progressive liver fibrosis leading to cirrhosis and liver cancer. The mechanism for this remains unclear but hepatocyte apoptosis is thought to play a major role. Hepatocyte apoptosis in human liver tissue was determined by immunohistochemistry for cytokeratin 18 (M30 CytoDEATH) and cleaved poly(ADP-ribose) polymerase (PARP). studies were performed with replication-defective recombinant adenoviruses expressing HCV proteins (rAdHCV) to study the effects of HCV on cell death in Huh7 cells, primary mouse hepatocytes (PMoHs) and primary human hepatocytes (PHHs). Cell viability and apoptosis were studied using crystal violet assays and Western blots probed for cleaved caspase-3 and cleaved PARP, with and without treatment with the pan-caspase inhibitor Q-VD-OPh and necrostatin-1. Liver tissue of HCV-infected patients expressed elevated levels of apoptotic markers compared with HCV-negative patients. rAdHCV infection reduced cell viability compared with uninfected controls and cells infected with control virus (rAdGFP). Huh7, PMoHs and PHHs infected with rAdHCV showed significantly increased levels of apoptotic markers compared with uninfected controls and rAdGFP-infected cells. In rAdHCV-infected Huh7, treatment with Q-VD-OPh and necrostatin-1 both improved cell viability. Q-VD-Oph also reduced cleaved PARP in rAdHCV-infected Huh7 and PMoHs. Hepatocyte apoptosis is known to be increased in the livers of HCV-infected patients. HCV promoted cell death in primary and immortalized hepatocytes, and this was inhibited by Q-VD-OPh and necrostatin-1. These findings indicate that HCV-induced cell death occurs by both apoptosis and necroptosis, and provide new insights into the mechanisms of HCV-induced liver injury.

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2014-10-01
2019-11-22
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References

  1. Bantel H., Schulze-Osthoff K.. ( 2003;). Apoptosis in hepatitis C virus infection. . Cell Death Differ 10: (Suppl 1), S48–S58. [CrossRef][PubMed]
    [Google Scholar]
  2. Bantel H., Lügering A., Heidemann J., Volkmann X., Poremba C., Strassburg C. P., Manns M. P., Schulze-Osthoff K.. ( 2004;). Detection of apoptotic caspase activation in sera from patients with chronic HCV infection is associated with fibrotic liver injury. . Hepatology 40:, 1078–1087. [CrossRef][PubMed]
    [Google Scholar]
  3. Berg C. P., Schlosser S. F., Neukirchen D. K. H., Papadakis C., Gregor M., Wesselborg S., Stein G. M.. ( 2009;). Hepatitis C virus core protein induces apoptosis-like caspase independent cell death. . Virol J 6:, 213. [CrossRef][PubMed]
    [Google Scholar]
  4. Brown G. D.. ( 2008;). Sensing necrosis with Mincle. . Nat Immunol 9:, 1099–1100. [CrossRef][PubMed]
    [Google Scholar]
  5. Canbay A., Feldstein A., Baskin-Bey E., Bronk S. F., Gores G. J.. ( 2004;). The caspase inhibitor IDN-6556 attenuates hepatic injury and fibrosis in the bile duct ligated mouse. . J Pharmacol Exp Ther 308:, 1191–1196. [CrossRef][PubMed]
    [Google Scholar]
  6. Chiou H.-L., Hsieh Y.-S., Hsieh M.-R., Chen T.-Y.. ( 2006;). HCV E2 may induce apoptosis of Huh-7 cells via a mitochondrial-related caspase pathway. . Biochem Biophys Res Commun 345:, 453–458. [CrossRef][PubMed]
    [Google Scholar]
  7. Chua B. Y., Johnson D., Tan A., Earnest-Silveira L., Sekiya T., Chin R., Torresi J., Jackson D. C.. ( 2012;). Hepatitis C VLPs delivered to dendritic cells by a TLR2 targeting lipopeptide results in enhanced antibody and cell-mediated responses. . PLoS ONE 7:, e47492. [CrossRef][PubMed]
    [Google Scholar]
  8. Ciccaglione A. R., Marcantonio C., Costantino A., Equestre M., Rapicetta M.. ( 2003;). Expression of HCV E1 protein in baculovirus-infected cells: effects on cell viability and apoptosis induction. . Intervirology 46:, 121–126. [CrossRef][PubMed]
    [Google Scholar]
  9. Declercq W., Vanden Berghe T., Vandenabeele P.. ( 2009;). RIP kinases at the crossroads of cell death and survival. . Cell 138:, 229–232. [CrossRef][PubMed]
    [Google Scholar]
  10. Deng L., Adachi T., Kitayama K., Bungyoku Y., Kitazawa S., Ishido S., Shoji I., Hotta H.. ( 2008;). Hepatitis C virus infection induces apoptosis through a Bax-triggered, mitochondrion-mediated, caspase 3-dependent pathway. . J Virol 82:, 10375–10385. [CrossRef][PubMed]
    [Google Scholar]
  11. Dore G. J., Law M., MacDonald M., Kaldor J. M.. ( 2003;). Epidemiology of hepatitis C virus infection in Australia. . J Clin Virol 26:, 171–184. [CrossRef][PubMed]
    [Google Scholar]
  12. Fabregat I., Roncero C., Fernández M.. ( 2007;). Survival and apoptosis: a dysregulated balance in liver cancer. . Liver Int 27:, 155–162. [CrossRef][PubMed]
    [Google Scholar]
  13. Fischer R., Baumert T., Blum H.-E.. ( 2007;). Hepatitis C virus infection and apoptosis. . World J Gastroenterol 13:, 4865–4872.[PubMed]
    [Google Scholar]
  14. Jost P. J., Grabow S., Gray D., McKenzie M. D., Nachbur U., Huang D. C. S., Bouillet P., Thomas H. E., Borner C.. & other authors ( 2009;). XIAP discriminates between type I and type II FAS-induced apoptosis. . Nature 460:, 1035–1039. [CrossRef][PubMed]
    [Google Scholar]
  15. Kawamura H., Govindarajan S., Aswad F., Machida K., Lai M. M. C., Sung V. M. H., Dennert G.. ( 2006;). HCV core expression in hepatocytes protects against autoimmune liver injury and promotes liver regeneration in mice. . Hepatology 44:, 936–944. [CrossRef][PubMed]
    [Google Scholar]
  16. Lan L., Gorke S., Rau S. J., Zeisel M. B., Hildt E., Himmelsbach K., Carvajal-Yepes M., Huber R., Wakita T.. & other authors ( 2008;). Hepatitis C virus infection sensitizes human hepatocytes to TRAIL-induced apoptosis in a caspase 9-dependent manner. . J Immunol 181:, 4926–4935. [CrossRef][PubMed]
    [Google Scholar]
  17. Lauber K., Blumenthal S. G., Waibel M., Wesselborg S.. ( 2004;). Clearance of apoptotic cells: getting rid of the corpses. . Mol Cell 14:, 277–287. [CrossRef][PubMed]
    [Google Scholar]
  18. Lavanchy D.. ( 2009;). The global burden of hepatitis C. . Liver Int 29: (Suppl 1), 74–81. [CrossRef][PubMed]
    [Google Scholar]
  19. Liang T. J., Heller T.. ( 2004;). Pathogenesis of hepatitis C-associated hepatocellular carcinoma. . Gastroenterology 127: (Suppl 1), S62–S71. [CrossRef][PubMed]
    [Google Scholar]
  20. Macdonald A., Harris M.. ( 2004;). Hepatitis C virus NS5A: tales of a promiscuous protein. . J Gen Virol 85:, 2485–2502. [CrossRef][PubMed]
    [Google Scholar]
  21. Nomura-Takigawa Y., Nagano-Fujii M., Deng L., Kitazawa S., Ishido S., Sada K., Hotta H.. ( 2006;). Non-structural protein 4A of Hepatitis C virus accumulates on mitochondria and renders the cells prone to undergoing mitochondria-mediated apoptosis. . J Gen Virol 87:, 1935–1945. [CrossRef][PubMed]
    [Google Scholar]
  22. Ogata H., Kobayashi T., Chinen T., Takaki H., Sanada T., Minoda Y., Koga K., Takaesu G., Maehara Y.. & other authors ( 2006;). Deletion of the SOCS3 gene in liver parenchymal cells promotes hepatitis-induced hepatocarcinogenesis. . Gastroenterology 131:, 179–193. [CrossRef][PubMed]
    [Google Scholar]
  23. Paizis G., Tikellis C., Cooper M. E., Schembri J. M., Lew R. A., Smith A. I., Shaw T., Warner F. J., Zuilli A.. & other authors ( 2005;). Chronic liver injury in rats and humans upregulates the novel enzyme angiotensin converting enzyme 2. . Gut 54:, 1790–1796. [CrossRef][PubMed]
    [Google Scholar]
  24. Petersen S. L., Wang L., Yalcin-Chin A., Li L., Peyton M., Minna J., Harran P., Wang X.. ( 2007;). Autocrine TNFalpha signaling renders human cancer cells susceptible to Smac-mimetic-induced apoptosis. . Cancer Cell 12:, 445–456. [CrossRef][PubMed]
    [Google Scholar]
  25. Pianko S., Patella S., Ostapowicz G., Desmond P., Sievert W.. ( 2001;). Fas-mediated hepatocyte apoptosis is increased by hepatitis C virus infection and alcohol consumption, and may be associated with hepatic fibrosis: mechanisms of liver cell injury in chronic hepatitis C virus infection. . J Viral Hepat 8:, 406–413. [CrossRef][PubMed]
    [Google Scholar]
  26. Prikhod’ko E. A., Prikhod’ko G. G., Siegel R. M., Thompson P., Major M. E., Cohen J. I.. ( 2004;). The NS3 protein of hepatitis C virus induces caspase-8-mediated apoptosis independent of its protease or helicase activities. . Virology 329:, 53–67. [CrossRef][PubMed]
    [Google Scholar]
  27. Ruggieri A., Harada T., Matsuura Y., Miyamura T.. ( 1997;). Sensitization to Fas-mediated apoptosis by hepatitis C virus core protein. . Virology 229:, 68–76. [CrossRef][PubMed]
    [Google Scholar]
  28. Saito K., Meyer K., Warner R., Basu A., Ray R. B., Ray R.. ( 2006;). Hepatitis C virus core protein inhibits tumor necrosis factor α-mediated apoptosis by a protective effect involving cellular FLICE inhibitory protein. . J Virol 80:, 4372–4379. [CrossRef][PubMed]
    [Google Scholar]
  29. Schattenberg J. M., Galle P. R., Schuchmann M.. ( 2006;). Apoptosis in liver disease. . Liver Int 26:, 904–911. [CrossRef][PubMed]
    [Google Scholar]
  30. Shackel N. A., McGuinness P. H., Abbott C. A., Gorrell M. D., McCaughan G. W.. ( 2002;). Insights into the pathobiology of hepatitis C virus-associated cirrhosis: analysis of intrahepatic differential gene expression. . Am J Pathol 160:, 641–654. [CrossRef][PubMed]
    [Google Scholar]
  31. Shiffman M. L., Pockros P., McHutchison J. G., Schiff E. R., Morris M., Burgess G.. ( 2010;). Clinical trial: the efficacy and safety of oral PF-03491390, a pancaspase inhibitor – a randomized placebo-controlled study in patients with chronic hepatitis C. . Aliment Pharmacol Ther 31:, 969–978.[PubMed]
    [Google Scholar]
  32. Takehara T., Tatsumi T., Suzuki T., Rucker E. B. III, Hennighausen L., Jinushi M., Miyagi T., Kanazawa Y., Hayashi N.. ( 2004;). Hepatocyte-specific disruption of Bcl-xL leads to continuous hepatocyte apoptosis and liver fibrotic responses. . Gastroenterology 127:, 1189–1197. [CrossRef][PubMed]
    [Google Scholar]
  33. Taniguchi H., Kato N., Otsuka M., Goto T., Yoshida H., Shiratori Y., Omata M.. ( 2004;). Hepatitis C virus core protein upregulates transforming growth factor-beta 1 transcription. . J Med Virol 72:, 52–59. [CrossRef][PubMed]
    [Google Scholar]
  34. Teixeira R., Marcos L. A., Friedman S. L.. ( 2007;). Immunopathogenesis of hepatitis C virus infection and hepatic fibrosis: new insights into antifibrotic therapy in chronic hepatitis C. . Hepatol Res 37:, 579–595. [CrossRef][PubMed]
    [Google Scholar]
  35. Vandenabeele P., Galluzzi L., Vanden Berghe T., Kroemer G.. ( 2010;). Molecular mechanisms of necroptosis: an ordered cellular explosion. . Nat Rev Mol Cell Biol 11:, 700–714. [CrossRef][PubMed]
    [Google Scholar]
  36. Varfolomeev E., Blankenship J. W., Wayson S. M., Fedorova A. V., Kayagaki N., Garg P., Zobel K., Dynek J. N., Elliott L. O.. & other authors ( 2007;). IAP antagonists induce autoubiquitination of c-IAPs, NF-kappaB activation, and TNFalpha-dependent apoptosis. . Cell 131:, 669–681. [CrossRef][PubMed]
    [Google Scholar]
  37. Witek R. P., Stone W. C., Karaca F. G., Syn W.-K., Pereira T. A., Agboola K. M., Omenetti A., Jung Y., Teaberry V.. & other authors ( 2009;). Pan-caspase inhibitor VX-166 reduces fibrosis in an animal model of nonalcoholic steatohepatitis. . Hepatology 50:, 1421–1430. [CrossRef][PubMed]
    [Google Scholar]
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