1887

Abstract

Amphiregulin (AREG) is a ligand of the epidermal growth factor (EGF) receptor and may play a role in the development of cirrhosis and hepatocellular carcinoma in patients infected with hepatitis C virus (HCV). AREG showed an enhanced expression in HCV-infected human hepatoma cells according to gene array analysis. Therefore, we addressed the question about the role of AREG in HCV infection. AREG expression level was elevated in hepatoma cells containing a subgenomic HCV replicon or infected by HCV. Using a reporter assay, AREG promoter activity was found to be upregulated upon HCV infection. The enhanced AREG expression in hepatoma cells was partly caused by dsRNAs, HCV NS3 protein and autocrine stimulation. AREG was able to activate cellular signalling pathways including ERK, Akt and p38, promote cell proliferation, and protect cells from HCV-induced cell death. Further, knockdown of AREG expression increased the efficiency of HCV entry, as proven by HCV pseudoparticles reporter assay. However, the formation and release of infectious HCV particles were reduced by AREG silencing with a concomitant accumulation of intracellular HCV RNA pool, indicating that the assembly and release of HCV progeny may require AREG expression. Blocking the MAPK–ERK pathway by U0126 in Huh7.5.1 cells had a similar effect on HCV replication. In conclusion, HCV infection leads to an increase in AREG expression in hepatocytes. AREG expression is essential for efficient HCV assembly and virion release. Due to the activation of the cellular survival pathways, AREG may counteract HCV-induced apoptosis of infected hepatocytes and facilitate the development of liver cirrhosis and hepatocellular carcinoma.

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2011-10-01
2020-01-23
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References

  1. Baillo A. , Giroux C. , Ethier S. P. . ( 2011; ). Knock-down of amphiregulin inhibits cellular invasion in inflammatory breast cancer. . J Cell Physiol 00:, 00–00 (in press). [CrossRef] [PubMed]
    [Google Scholar]
  2. Berasain C. , García-Trevijano E. R. , Castillo J. , Erroba E. , Lee D. C. , Prieto J. , Avila M. A. . ( 2005a; ). Amphiregulin: an early trigger of liver regeneration in mice. . Gastroenterology 128:, 424–432. [CrossRef] [PubMed]
    [Google Scholar]
  3. Berasain C. , García-Trevijano E. R. , Castillo J. , Erroba E. , Santamaría M. , Lee D. C. , Prieto J. , Avila M. A. . ( 2005b; ). Novel role for amphiregulin in protection from liver injury. . J Biol Chem 280:, 19012–19020. [CrossRef] [PubMed]
    [Google Scholar]
  4. Berasain C. , Castillo J. , Perugorría M. J. , Prieto J. , Avila M. A. . ( 2007; ). Amphiregulin: a new growth factor in hepatocarcinogenesis. . Cancer Lett 254:, 30–41. [CrossRef] [PubMed]
    [Google Scholar]
  5. Berger K. L. , Cooper J. D. , Heaton N. S. , Yoon R. , Oakland T. E. , Jordan T. X. , Mateu G. , Grakoui A. , Randall G. . ( 2009; ). Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication. . Proc Natl Acad Sci U S A 106:, 7577–7582. [CrossRef] [PubMed]
    [Google Scholar]
  6. Berquin I. M. , Dziubinski M. L. , Nolan G. P. , Ethier S. P. . ( 2001; ). A functional screen for genes inducing epidermal growth factor autonomy of human mammary epithelial cells confirms the role of amphiregulin. . Oncogene 20:, 4019–4028. [CrossRef] [PubMed]
    [Google Scholar]
  7. Blackham S. , Baillie A. , Al-Hababi F. , Remlinger K. , You S. , Hamatake R. , McGarvey M. J. . ( 2010; ). Gene expression profiling indicates the roles of host oxidative stress, apoptosis, lipid metabolism, and intracellular transport genes in the replication of hepatitis C virus. . J Virol 84:, 5404–5414. [CrossRef] [PubMed]
    [Google Scholar]
  8. Borawski J. , Troke P. , Puyang X. , Gibaja V. , Zhao S. , Mickanin C. , Leighton-Davies J. , Wilson C. J. , Myer V. et al. & other authors ( 2009; ). Class III phosphatidylinositol 4-kinase alpha and beta are novel host factor regulators of hepatitis C virus replication. . J Virol 83:, 10058–10074. [CrossRef] [PubMed]
    [Google Scholar]
  9. Bordoli M. R. , Stiehl D. P. , Borsig L. , Kristiansen G. , Hausladen S. , Schraml P. , Wenger R. H. , Camenisch G. . ( 2010; ). Prolyl-4-hydroxylase PHD2- and hypoxia-inducible factor 2-dependent regulation of amphiregulin contributes to breast tumorigenesis. . Oncogene 30:, 548–560.[PubMed] [CrossRef]
    [Google Scholar]
  10. Brenndörfer E. D. , Karthe J. , Frelin L. , Cebula P. , Erhardt A. , Schulte am Esch J. , Hengel H. , Bartenschlager R. , Sällberg M. et al. & other authors ( 2009; ). Nonstructural 3/4A protease of hepatitis C virus activates epithelial growth factor-induced signal transduction by cleavage of the T-cell protein tyrosine phosphatase. . Hepatology 49:, 1810–1820. [CrossRef] [PubMed]
    [Google Scholar]
  11. Burdette D. , Olivarez M. , Waris G. . ( 2010; ). Activation of transcription factor Nrf2 by hepatitis C virus induces the cell-survival pathway. . J Gen Virol 91:, 681–690. [CrossRef] [PubMed]
    [Google Scholar]
  12. Castillo J. , Erroba E. , Perugorría M. J. , Santamaría M. , Lee D. C. , Prieto J. , Avila M. A. , Berasain C. . ( 2006; ). Amphiregulin contributes to the transformed phenotype of human hepatocellular carcinoma cells. . Cancer Res 66:, 6129–6138. [CrossRef] [PubMed]
    [Google Scholar]
  13. Castillo J. , Goñi S. , Latasa M. U. , Perugorría M. J. , Calvo A. , Muntané J. , Bioulac-Sage P. , Balabaud C. , Prieto J. et al. & other authors ( 2009; ). Amphiregulin induces the alternative splicing of p73 into its oncogenic isoform DeltaEx2p73 in human hepatocellular tumors. . Gastroenterology 137:, 1805–1815, e1–e4. [CrossRef] [PubMed]
    [Google Scholar]
  14. Coller K. E. , Berger K. L. , Heaton N. S. , Cooper J. D. , Yoon R. , Randall G. . ( 2009; ). RNA interference and single particle tracking analysis of hepatitis C virus endocytosis. . PLoS Pathog 5:, e1000702. [CrossRef] [PubMed]
    [Google Scholar]
  15. Connor R. I. , Chen B. K. , Choe S. , Landau N. R. . ( 1995; ). Vpr is required for efficient replication of human immunodeficiency virus type-1 in mononuclear phagocytes. . Virology 206:, 935–944. [CrossRef] [PubMed]
    [Google Scholar]
  16. 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]
  17. Diamond D. L. , Jacobs J. M. , Paeper B. , Proll S. C. , Gritsenko M. A. , Carithers R. L. Jr , Larson A. M. , Yeh M. M. , Camp D. G. II et al. & other authors ( 2007; ). Proteomic profiling of human liver biopsies: hepatitis C virus-induced fibrosis and mitochondrial dysfunction. . Hepatology 46:, 649–657. [CrossRef] [PubMed]
    [Google Scholar]
  18. Fang C. , Yi Z. , Liu F. , Lan S. , Wang J. , Lu H. , Yang P. , Yuan Z. . ( 2006a; ). Proteome analysis of human liver carcinoma Huh7 cells harboring hepatitis C virus subgenomic replicon. . Proteomics 6:, 519–527. [CrossRef] [PubMed]
    [Google Scholar]
  19. Fang X. , Zeisel M. B. , Wilpert J. , Gissler B. , Thimme R. , Kreutz C. , Maiwald T. , Timmer J. , Kern W. V. et al. & other authors ( 2006b; ). Host cell responses induced by hepatitis C virus binding. . Hepatology 43:, 1326–1336. [CrossRef] [PubMed]
    [Google Scholar]
  20. Han Q. , Xu C. , Wu C. , Zhu W. , Yang R. , Chen X. . ( 2009; ). Compensatory mutations in NS3 and NS5A proteins enhance the virus production capability of hepatitis C reporter virus. . Virus Res 145:, 63–73. [CrossRef] [PubMed]
    [Google Scholar]
  21. Joyce M. A. , Walters K. A. , Lamb S. E. , Yeh M. M. , Zhu L. F. , Kneteman N. , Doyle J. S. , Katze M. G. , Tyrrell D. L. . ( 2009; ). HCV induces oxidative and ER stress, and sensitizes infected cells to apoptosis in SCID/Alb-uPA mice. . PLoS Pathog 5:, e1000291. [CrossRef] [PubMed]
    [Google Scholar]
  22. Kasina S. , Scherle P. A. , Hall C. L. , Macoska J. A. . ( 2009; ). ADAM-mediated amphiregulin shedding and EGFR transactivation. . Cell Prolif 42:, 799–812. [CrossRef] [PubMed]
    [Google Scholar]
  23. Lai C. K. , Jeng K. S. , Machida K. , Lai M. M. . ( 2010; ). Hepatitis C virus egress and release depend on endosomal trafficking of core protein. . J Virol 84:, 11590–11598. [CrossRef] [PubMed]
    [Google Scholar]
  24. Lamber E. P. , Horwitz A. A. , Parvin J. D. . ( 2010; ). BRCA1 represses amphiregulin gene expression. . Cancer Res 70:, 996–1005. [CrossRef] [PubMed]
    [Google Scholar]
  25. Li Q. , Brass A. L. , Ng A. , Hu Z. , Xavier R. J. , Liang T. J. , Elledge S. J. . ( 2009; ). A genome-wide genetic screen for host factors required for hepatitis C virus propagation. . Proc Natl Acad Sci U S A 106:, 16410–16415. [CrossRef] [PubMed]
    [Google Scholar]
  26. Li R. , Qin Y. , He Y. , Tao W. , Zhang N. , Tsai C. , Zhou P. , Zhong J. . ( 2011; ). Production of hepatitis C virus lacking the envelope-encoding genes for single-cycle infection by providing homologous envelope proteins or vesicular stomatitis virus glycoproteins in trans. . J Virol 85:, 2138–2147. [CrossRef] [PubMed]
    [Google Scholar]
  27. Lindenbach B. D. . ( 2009; ). Measuring HCV infectivity produced in cell culture and in vivo . . Methods Mol Biol 510:, 329–336. [CrossRef] [PubMed]
    [Google Scholar]
  28. Lupberger J. , Zeisel M. B. , Xiao F. , Thumann C. , Fofana I. , Zona L. , Davis C. , Mee C. J. , Turek M. et al. & other authors ( 2011; ). EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy. . Nat Med 17:, 589–595. [CrossRef] [PubMed]
    [Google Scholar]
  29. Mannová P. , Beretta L. . ( 2005; ). Activation of the N-Ras-PI3K-Akt-mTOR pathway by hepatitis C virus: control of cell survival and viral replication. . J Virol 79:, 8742–8749. [CrossRef] [PubMed]
    [Google Scholar]
  30. McBryan J. , Howlin J. , Napoletano S. , Martin F. . ( 2008; ). Amphiregulin: role in mammary gland development and breast cancer. . J Mammary Gland Biol Neoplasia 13:, 159–169. [CrossRef] [PubMed]
    [Google Scholar]
  31. Ng T. I. , Mo H. , Pilot-Matias T. , He Y. , Koev G. , Krishnan P. , Mondal R. , Pithawalla R. , He W. et al. & other authors ( 2007; ). Identification of host genes involved in hepatitis C virus replication by small interfering RNA technology. . Hepatology 45:, 1413–1421. [CrossRef] [PubMed]
    [Google Scholar]
  32. Pardo-Saganta A. , Latasa M. U. , Castillo J. , Alvarez-Asiain L. , Perugorría M. J. , Sarobe P. , Rodriguez-Ortigosa C. M. , Prieto J. , Berasain C. , Santamaría M. . ( 2009; ). The epidermal growth factor receptor ligand amphiregulin is a negative regulator of hepatic acute-phase gene expression. . J Hepatol 51:, 1010–1020. [CrossRef] [PubMed]
    [Google Scholar]
  33. Perugorria M. J. , Latasa M. U. , Nicou A. , Cartagena-Lirola H. , Castillo J. , Goñi S. , Vespasiani-Gentilucci U. , Zagami M. G. , Lotersztajn S. et al. & other authors ( 2008; ). The epidermal growth factor receptor ligand amphiregulin participates in the development of mouse liver fibrosis. . Hepatology 48:, 1251–1261. [CrossRef] [PubMed]
    [Google Scholar]
  34. Plowman G. D. , Green J. M. , McDonald V. L. , Neubauer M. G. , Disteche C. M. , Todaro G. J. , Shoyab M. . ( 1990; ). The amphiregulin gene encodes a novel epidermal growth factor-related protein with tumor-inhibitory activity. . Mol Cell Biol 10:, 1969–1981.[PubMed]
    [Google Scholar]
  35. Randall G. , Panis M. , Cooper J. D. , Tellinghuisen T. L. , Sukhodolets K. E. , Pfeffer S. , Landthaler M. , Landgraf P. , Kan S. et al. & other authors ( 2007; ). Cellular cofactors affecting hepatitis C virus infection and replication. . Proc Natl Acad Sci U S A 104:, 12884–12889. [CrossRef] [PubMed]
    [Google Scholar]
  36. Tai A. W. , Benita Y. , Peng L. F. , Kim S. S. , Sakamoto N. , Xavier R. J. , Chung R. T. . ( 2009; ). A functional genomic screen identifies cellular cofactors of hepatitis C virus replication. . Cell Host Microbe 5:, 298–307. [CrossRef] [PubMed]
    [Google Scholar]
  37. Vaillancourt F. H. , Pilote L. , Cartier M. , Lippens J. , Liuzzi M. , Bethell R. C. , Cordingley M. G. , Kukolj G. . ( 2009; ). Identification of a lipid kinase as a host factor involved in hepatitis C virus RNA replication. . Virology 387:, 5–10. [CrossRef] [PubMed]
    [Google Scholar]
  38. Walters K. A. , Syder A. J. , Lederer S. L. , Diamond D. L. , Paeper B. , Rice C. M. , Katze M. G. . ( 2009; ). Genomic analysis reveals a potential role for cell cycle perturbation in HCV-mediated apoptosis of cultured hepatocytes. . PLoS Pathog 5:, e1000269. [CrossRef] [PubMed]
    [Google Scholar]
  39. Wang X. , Masri S. , Phung S. , Chen S. . ( 2008; ). The role of amphiregulin in exemestane-resistant breast cancer cells: evidence of an autocrine loop. . Cancer Res 68:, 2259–2265. [CrossRef] [PubMed]
    [Google Scholar]
  40. Watanabe T. , Kobunai T. , Yamamoto Y. , Kanazawa T. , Konishi T. , Tanaka T. , Matsuda K. , Ishihara S. , Nozawa K. et al. & other authors ( 2010; ). Prediction of liver metastasis after colorectal cancer using reverse transcription-polymerase chain reaction analysis of 10 genes. . Eur J Cancer 46:, 2119–2126. [CrossRef] [PubMed]
    [Google Scholar]
  41. Woodhouse S. D. , Narayan R. , Latham S. , Lee S. , Antrobus R. , Gangadharan B. , Luo S. , Schroth G. P. , Klenerman P. , Zitzmann N. . ( 2010; ). Transcriptome sequencing, microarray, and proteomic analyses reveal cellular and metabolic impact of hepatitis C virus infection in vitro . . Hepatology 52:, 443–453. [CrossRef] [PubMed]
    [Google Scholar]
  42. Yotsumoto F. , Yagi H. , Suzuki S. O. , Oki E. , Tsujioka H. , Hachisuga T. , Sonoda K. , Kawarabayashi T. , Mekada E. , Miyamoto S. . ( 2008; ). Validation of HB-EGF and amphiregulin as targets for human cancer therapy. . Biochem Biophys Res Commun 365:, 555–561. [CrossRef] [PubMed]
    [Google Scholar]
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Supplements

vol. , part 10, pp. 2237 - 2248

IFN-α treatment does not change AREG expression in Huh7-Lunet cells

Sendai virus infection upregulates AREG mRNA levels in Huh7-Lunet cells but not in Huh7.5.1 cells

Poly IC is not able to activate AREG promoter in Huh7.5.1 cells

Stimulation of AREG expression by rhAREG in Con1 cells

Treatment with exogenous rhAREG did not influence HCV replication in Con1 cells

Knockdown of AREG expression affects HCV induced activation of cell signalling pathway

Primers used in the experiments [Single PDF file](373 KB)

 



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