1887

Abstract

The hepatitis B virus-encoded X antigen (HBxAg) may contribute to the development of liver cancer, in part, by stimulating the growth and survival of infected cells in the face of ongoing immune responses. Given that the Fas ligand/receptor system contributes to the pathogenesis of chronic hepatitis B, experiments were designed to test the hypothesis that HBxAg mediates resistance of liver cells to anti-Fas killing. Accordingly, when HBxAg was introduced into HepG2 cells, it rendered these cells partially resistant to killing by anti-Fas. In HepG2 cells replicating virus, protection against anti-Fas killing was also observed, but to a lesser extent. Survival correlated with the activation of nuclear factor kappa B (NF-κB) by HBxAg. Sensitivity to anti-Fas was observed in control cells, and was re-established in HepG2X cells stably transfected with the dominant negative inhibitor of NF-κB, IκBα. HBxAg activation of NF-κB was also associated with decreased levels of endogenous IκBα mRNA. Hence, HBxAg stimulation of NF-κB promotes the survival of liver cells against Fas killing. This may contribute to the persistence of infected hepatocytes during chronic infection.

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2001-01-01
2019-10-16
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References

  1. Ando, K., Guidotti, L. G., Wirth, S., Ishikawa, T., Missale, G., Moriyama, T., Schreiber, R. D., Schlicht, H. J. & Chisari, F. V. ( 1994; ). Class I-restricted cytotoxic T lymphocytes are directly cytopathic for their target cells. Journal of lmmunology 152, 3245-3253.
    [Google Scholar]
  2. Arsura, M., FitzGerald, M. J., Fausto, N. & Sonenshein, G. E. ( 1997; ). Nuclear factor-κB/Rel blocks transforming growth factor β1-induced apoptosis of murine hepatocyte cell lines. Cell Growth & Differentiation 8, 1049-1059.
    [Google Scholar]
  3. Ausubel, F., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K. (1991). Current Protocols in Molecular Biology, vol. 1. New York: John Wiley.
  4. Baeuerle, P. A. ( 1991; ). The inducible transcription activator NF-κB: regulation by distinct protein subunits. Biochimica et Biophysica Acta 1072, 63-80.
    [Google Scholar]
  5. Balsano, C., Avantaggiati, M. L., Natoli, G., De, M., Will, H., Perricaudet, M. & Levrero, M. ( 1991; ). Full-length and truncated versions of the hepatitis B virus (HBV) X protein (pX) transactivate the c-myc protooncogene at the transcriptional level. Biochemical and Biophysical Research Communications 176, 985-992.[CrossRef]
    [Google Scholar]
  6. Beg, A. A. & Baltimore, D. ( 1996; ). An essential role for NF-κB in preventing TNF-α-induced cell death. Science 274, 782-784.[CrossRef]
    [Google Scholar]
  7. Beg, A. A., Sha, W. C., Bronson, R. T., Ghosh, S. & Baltimore, D. ( 1995; ). Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-κB. Nature 376, 167-170.[CrossRef]
    [Google Scholar]
  8. Bellas, R. E., FitzGerald, M. J., Fausto, N. & Sonenshein, G. E. ( 1997; ). Inhibition of NF-κB activity induces apoptosis in murine hepatocytes. American Journal of Pathology 151, 891-896.
    [Google Scholar]
  9. Benn, J. & Schneider, R. J. ( 1995; ). HBV HBx protein deregulates cell cycle checkpoint controls. Proceedings of the National Academy of Sciences, USA 92, 11215-11219.[CrossRef]
    [Google Scholar]
  10. Chirillo, P., Falco, M., Puri, P. L., Artini, M., Balsano, C., Levrero, M. & Natoli, G. ( 1996; ). Hepatitis B virus pX activates NF-κB dependent transcription through a Raf-independent pathway. Journal of Virology 70, 641-646.
    [Google Scholar]
  11. Colgrove, R., Simon, G. & Ganem, D. ( 1989; ). Transcriptional activation of homologous and heterologous genes by the HBV X gene product in cells permissive for viral replication. Journal of Virology 63, 4019-4026.
    [Google Scholar]
  12. Diamantis, I. D., McGandy, C. E., Chen, T. J., Liaw, Y. F., Gudat, F. & Bianchi, L. ( 1992; ). Hepatitis B X-gene expression in hepatocellular carcinoma. Journal of Hepatology 15, 400-403.[CrossRef]
    [Google Scholar]
  13. Doria, M., Klein, N., Lucito, R. & Schneider, R. J. ( 1995; ). The hepatitis B virus HBx protein is a dual specificity cytoplasmic activator of Ras and nuclear activator of transcription factors. EMBO Journal 14, 4747-4757.
    [Google Scholar]
  14. Duan, L.-X., Zhu, M.-H., Bagasra, O. & Pomerantz, R. J. ( 1995; ). Intracellular immunization against HIV-1 infection of human T lymphocytes: utility of anti-Rev single-chain variable fragments. Human Gene Therapy 6, 1561-1573.[CrossRef]
    [Google Scholar]
  15. Dudley, E., Hornung, F., Zheng, F., Scherer, D., Ballard, D. & Lenardo, M. ( 1999; ). NF-kappaB regulates Fas/APO-1/CD95- and TCR-mediated apoptosis of T lymphocytes. European Journal of Immunology 29, 878-886.[CrossRef]
    [Google Scholar]
  16. Elmore, L. W., Hancock, A. R., Chang, S. F., Wang, X. W., Chang, S., Callahan, C. P., Geller, D. A., Will, H. & Harris, C. C. ( 1997; ). Hepatitis B virus X protein and p53 tumor suppressor interactions in the modulation of apoptosis. Proceedings of the National Academy of Sciences, USA 94, 14707-14712.[CrossRef]
    [Google Scholar]
  17. Feitelson, M. A. & Duan, L.-X. ( 1997; ). Hepatitis B virus x antigen in the pathogenesis of chronic infections and the development of hepatocellular carcinoma. American Journal of Pathology 150, 1141-1157.
    [Google Scholar]
  18. Feitelson, M. A., Lega, L., Duan, L.-X. & Clayton, M. (1993a). Characteristics of woodchuck hepatitis X antigen in the livers and sera from chronically infected animals. Journal of Hepatology 17 (Suppl. 3), S24–S34.
  19. Feitelson, M. A., Zhu, M., Duan, L.-X. & London, W. T. ( 1993b; ). HBxAg and p53 are associated in vitro and in liver tissues from patients with primary hepatocellular carcinoma. Oncogene 8, 1109-1117.
    [Google Scholar]
  20. Feitelson, M. A., Reis, H., Pan, J., Lian, Z., Fang, J., Liu, J., Zhu, X., Zhu, M. & Sun, B. ( 1999; ). Abrogation of negative growth regulatory pathways by hepatitis B virus encoded X antigen in the development of hepatocellular carcinoma. In Normal and Malignant Liver Cell Growth, chapter 15 , pp. 156-170. Edited by W. E. Fleig. Lancaster, UK:Kluwer Academic Publishers.
  21. Galle, P. R. & Krammer, P. H. ( 1998; ). CD95-induced apoptosis in human liver disease. Seminars in Liver Disease 18, 141-151.[CrossRef]
    [Google Scholar]
  22. Galle, P. R., Hofmann, W. J., Walczak, H., Schaller, H., Otto, G. & Stremmel, N. ( 1995; ). Involvement of the Apo-1/Fas (CD95) receptor and ligand in liver damage. Journal of Experimental Medicine 182, 1223-1230.[CrossRef]
    [Google Scholar]
  23. Gorman, C., Moffat, L. F. & Howard, B. H. ( 1992; ). Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Molecular and Cellular Biology 2, 1044-1051.
    [Google Scholar]
  24. Gottlob, K., Fulco, M., Levrero, M. & Graessmann, A. ( 1998; ). The hepatitis B virus HBx protein inhibits caspase 3 activity. Journal of Biological Chemistry 273, 33347-33353.[CrossRef]
    [Google Scholar]
  25. Grilli, M., Chiu, J.-S. & Lenardo, M. J. ( 1993; ). NF-κB and Rel: participants in a multiform transcriptional regulatory system. International Review of Cytology 143, 1-62.
    [Google Scholar]
  26. Haskill, S., Beg, A. A., Tompkins, S. M., Morris, J. S., Yurochko, A. D., Sampson-Johannes, A., Mondal, K., Ralph, P. & Baldwin, A. S.Jr ( 1991; ). Characterization of an immediate-early gene induced in adherent monocytes that encodes IκB-like activity. Cell 65, 1281-1289.[CrossRef]
    [Google Scholar]
  27. Henkler, F. & Koshy, R. ( 1996; ). Hepatitis B virus transcriptional activators: mechanisms and possible role in oncogenesis. Journal of Viral Hepatitis 3, 109-121.[CrossRef]
    [Google Scholar]
  28. Hohne, M., Schaefer, S., Seifer, M., Feitelson, M., Paul, D. & Gerlich, W. H. ( 1990; ). Malignant transformation of immortalized hepatocytes by hepatitis B virus DNA. EMBO Journal 9, 1137-1145.
    [Google Scholar]
  29. Kasibhatla, S., Genestier, L. & Green, D. R. ( 1999; ). Regulation of fas-ligand expression during activation-induced cell death in T lymphocytes via nuclear factor kappaB. Journal of Biological Chemistry 274, 987-992.[CrossRef]
    [Google Scholar]
  30. Kekule, A. S., Lauer, U., Weiss, L., Luber, B. & Hofschneider, P. H. ( 1993; ). HBV transactivator HBx uses a tumour promoter signalling pathway. Nature 361, 742-745.[CrossRef]
    [Google Scholar]
  31. Kim, C. M., Koike, K., Saito, I., Miyamura, T. & Jay, G. ( 1991; ). HBx gene of HBV induces liver cancer in transgenic mice. Nature 351, 317-320.[CrossRef]
    [Google Scholar]
  32. Kim, H., Lee, H. & Yun, Y. ( 1998; ). X-gene induces apoptosis in liver cells. Journal of Biological Chemistry 273, 381-385.[CrossRef]
    [Google Scholar]
  33. Koike, K., Moriya, K., Iino, S., Yotsuyanagi, H., Endo, Y., Miyamura, T. & Kurokawa, K. ( 1994; ). High-level expression of hepatitis B virus HBx gene and hepatocarcinogenesis in transgenic mice. Hepatology 19, 810-819.[CrossRef]
    [Google Scholar]
  34. Lau, J. Y. N., Xie, X., Lai, M. M. C. & Wu, P. C. ( 1998; ). Apoptosis and viral hepatitis. Seminars in Liver Disease 18, 169-176.[CrossRef]
    [Google Scholar]
  35. Lee, T. H., Elledge, S. J. & Butel, J. S. ( 1995; ). HBV X protein interacts with a probable cellular DNA repair protein. Journal of Virology 69, 1107-1114.
    [Google Scholar]
  36. Lee, Y. I., Lee, S., Lee, Y., Bong, Y. S., Hyun, S. W., Yoo, Y. D., Kim, S. J., Kim, Y. W. & Poo, H. R. ( 1998; ). The human hepatitis B virus transactivator X gene product regulates Sp1 mediated transcription of an insulin-like growth factor II promoter 4. Oncogene 16, 2367-2380.[CrossRef]
    [Google Scholar]
  37. Li, Q., Antwerp, D. V., Mercurio, F., Lee, K.-F. & Verma, I. M. ( 1999; ). Severe liver degeneration in mice lacking the IκB kinase 2 gene. Science 284, 321-325.[CrossRef]
    [Google Scholar]
  38. Lian, Z., Pan, J., Liu, J., Zhu, M., Arbuthnot, P., Kew, M. C. & Feitelson, M. A. ( 1999; ). The translation initiation factor, SUI1, may be a target of hepatitis B x antigen in hepatocarcinogenesis. Oncogene 18, 1677-1687.[CrossRef]
    [Google Scholar]
  39. Lin, B., Williams-Skipp, C., Tao, Y., Schleicher, M. S., Cano, L. L., Duke, R. C. & Scheinman, R. I. ( 1999; ). NF-kappaB functions as both a proapoptotic and antiapoptotic regulatory factor within a single cell type. Cell Death & Differentiation 6, 570-582.[CrossRef]
    [Google Scholar]
  40. Liou, H. C. & Baltimore, D. ( 1993; ). Regulation of the NF-κB/rel transcription factor and IκB inhibitor system. Current Opinion in Cell Biology 5, 477-487.[CrossRef]
    [Google Scholar]
  41. Liu, Z.-G., Hsu, H., Goeddel, D. V. & Karin, M. ( 1996; ). Dissection of TNF receptor I effector function: JNK activation is not linked to apoptosis while NF-κB activation prevents cell death. Cell 87, 565-576.[CrossRef]
    [Google Scholar]
  42. Mahe, Y., Mukaida, N., Kuno, K., Akiyama, M., Ikeda, N., Matsushima, K. & Murakami, S. ( 1991; ). Hepatitis B virus X protein transactivates human interleukin-8 gene through acting on nuclear factor κB and CCAAT enhancer binding protein-like cis-elements. Journal of Biological Chemistry 266, 13759-13763.
    [Google Scholar]
  43. Marusawa, H., Hijikata, M., Chiba, T. & Shimotohno, K. ( 1999; ). Hepatitis C virus core protein inhibits Fas- and tumor necrosis factor alpha-mediated apoptosis via NF-kappaB activation. Journal of Virology 73, 4713-4720.
    [Google Scholar]
  44. May, M. J. & Ghosh, S. ( 1998; ). Signal transduction through NF-κB. Immunology Today 19, 80-88.[CrossRef]
    [Google Scholar]
  45. Menzo, S., Clementi, M., Alfani, E., Bagnarelli, P., Iacovacci, S., Manzin, A., Dandri, M., Natoli, G., Levrero, M. & Carloni, G. ( 1993; ). Trans-activation of epidermal growth factor receptor gene by the hepatitis B virus X-gene product. Virology 196, 878-882.[CrossRef]
    [Google Scholar]
  46. Miller, A. D. & Buttimore, C. ( 1986; ). Redesign of retrovirus packaging cell line to avoid recombination leading to helper virus formation. Molecular and Cellular Biology 6, 2895-2902.
    [Google Scholar]
  47. Miller, A. D. & Rosman, G. J. ( 1989; ). Improved retroviral vectors for gene transfer and expression. BioTechniques 7, 980-990.
    [Google Scholar]
  48. Mochizuki, K., Hayashi, N., Hiramatsu, N., Katayama, K., Kawanishi, Y., Kasahara, A., Fusamoto, H. & Kamada, T. ( 1996; ). Fas antigen expression in liver tissues of patients with chronic hepatitis B. Journal of Hepatology 24, 1-7.
    [Google Scholar]
  49. Nakamoto, Y., Guidotti, L. G., Pasquetto, V., Schreiber, R. D. & Chisari, F. V. ( 1997; ). Differential target cell sensitivity to CTL-activated death pathways in hepatitis B virus transgenic mice. Journal of Immunology 158, 5692-5697.
    [Google Scholar]
  50. Nakatake, H., Chisaka, O., Yamamoto, S., Matsubara, K. & Koshy, R. ( 1993; ). Effect of X protein on transactivation of HBV promoters and on viral replication. Virology 195, 305-314.[CrossRef]
    [Google Scholar]
  51. Natoli, G., Avantaggiati, M. L., Chirillo, P., Costanzo, A., Artini, M., Balsano, C. & Levrero, M. ( 1994a; ). Induction of the DNA-binding activity of c-Jun/c-Fos heterodimers by the hepatitis B virus transactivator pX. Molecular and Cellular Biology 14, 989-998.
    [Google Scholar]
  52. Natoli, G., Avantaggiati, M. L., Chirillo, P., Puri, P. L., Ianni, A., Balsano, C. & Levrero, M. ( 1994b; ). Ras- and Raf-dependant activation of c-jun transcriptional activity by the hepatitis B virus transactivator pX. Oncogene 9, 2837-2843.
    [Google Scholar]
  53. Ogasawara, J., Watanabe-Fukunage, R., Adachi, M., Matsuzawa, A., Kasugai, T., Kitamura, Y., Itoh, N., Suda, T. & Nagata, S. ( 1993; ). Lethal effect of the anti-Fas antibody in mice. Nature 364, 806-809.[CrossRef]
    [Google Scholar]
  54. Paterlini, P., Poussin, K., Kew, M., Franco, D. & Brechot, C. ( 1995; ). Selective accumulation of the X transcript of hepatitis B virus in patients negative for hepatitis B surface antigen with hepatocellular carcinoma. Hepatology 21, 313-321.
    [Google Scholar]
  55. Ramachandra, S. & Studzinski, G. P. (1995). Morphological and biochemical criteria of apoptosis. In Cell Growth and Apoptosis. A Practical Approach, chapter 7. Edited by G. P. Studzinski. New York: Oxford University Press.
  56. Rouquet, N., Allemand, I., Molina, T., Bennoun, M., Briand, P. & Joulin, V. ( 1995; ). Fas-dependent apoptosis is impaired by SV40-T antigen in transgenic liver. Oncogene 11, 1061-1067.
    [Google Scholar]
  57. Rouvier, E., Luciani, M. F. & Golstein, P. ( 1993; ). Fas involvement on Ca2+-independent T cell-mediated cytotoxicity. Journal of Experimental Medicine 177, 195-200.[CrossRef]
    [Google Scholar]
  58. Seifer, M., Hohne, M., Schaefer, S. & Gerlich, W. H. ( 1991; ). In vitro tumorigenicity of hepatitis B virus DNA and HBx protein. Journal of Hepatology 13, S61-S65.
    [Google Scholar]
  59. Sells, M. A., Chen, M. L. & Acs, G. ( 1987; ). Production of hepatitis B virus particles in HepG2 cells transfected with cloned hepatitis B virus DNA. Proceedings of the National Academy of Sciences, USA 84, 1005-1009.[CrossRef]
    [Google Scholar]
  60. Sells, M. A., Zelent, A. Z., Shvartsman, M. & Acs, G. ( 1988; ). Replicative intermediates of hepatitis B virus in HepG2 cells that produce infectious virions. Journal of Virology 62, 2836-2844.
    [Google Scholar]
  61. Shin, E. C., Shin, J. S., Park, J. H., Kim, H. & Kim, S. J. ( 1999; ). Expression of fas ligand in human hepatoma cell lines: role of hepatitis B virus (HBX) in induction of Fas ligand. International Journal of Cancer 82, 587-591.[CrossRef]
    [Google Scholar]
  62. Shirakata, Y., Kawada, M., Fujiki, Y., Sano, H., Oda, M., Yaginuma, K., Kobayashi, M. & Koike, K. ( 1989; ). The X gene of hepatitis B virus induced growth stimulation and tumorigenic transformation of mouse NIH3T3 cells. Japanese Journal of Cancer Research 80, 617-621.[CrossRef]
    [Google Scholar]
  63. Siddiqui, A., Gaynor, R., Srinivasan, A., Mapoles, J. & Farr, R. W. ( 1989; ). Trans-activation of viral enhancer including the long terminal repeat of HIV by the HBV X protein. Virology 169, 479-484.[CrossRef]
    [Google Scholar]
  64. Strand, S., Hofmann, W. J., Hug, H., Muller, M., Otto, G. & Strand, D. ( 1996; ). Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand expressing tumor cells – a mechanism of immune evasion? Nature Medicine 2, 1361-1367.[CrossRef]
    [Google Scholar]
  65. Su, F. & Schneider, R. J. ( 1996; ). HBV HBx protein activates transcription factor NF-κB by acting on multiple cytoplasmic inhibitors of rel-related proteins. Journal of Virology 70, 4558-4566.
    [Google Scholar]
  66. Suda, T., Takahasi, T., Golstein, P. & Nagata, S. ( 1993; ). Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell 75, 1169-1178.[CrossRef]
    [Google Scholar]
  67. Terradillos, O., Pollicino, T., Lecoeur, H., Tripodi, M., Gougeon, M.-L., Tiollais, P. & Buendia, M. A. ( 1998; ). p53-independent apoptotic effects of the hepatitis B virus HBx protein in vivo and in vitro. Oncogene 17, 2115-2123.[CrossRef]
    [Google Scholar]
  68. Tiollais, P., Pourcel, C. & Dejean, A. ( 1985; ). The hepatitis B virus. Nature 317, 489-495.[CrossRef]
    [Google Scholar]
  69. Truant, R., Antunovic, J., Greenblatt, J., Prives, C. & Cromlish, J. A. ( 1995; ). Direct interaction of the HBV HBx protein with p53 response element-directed transactivation. Journal of Virology 69, 1851-1859.
    [Google Scholar]
  70. Van Antwerp, D. J., Martin, S. J., Kafri, T., Green, D. R. & Verma, I. M. ( 1996; ). Suppression of TNF-α-induced apoptosis by NF-κB. Science 274, 787-789.[CrossRef]
    [Google Scholar]
  71. Wang, W., London, W. T. & Feitelson, M. A. ( 1991a; ). HBxAg in HBV carrier patients with liver cancer. Cancer Research 51, 4971-4977.
    [Google Scholar]
  72. Wang, W., London, W. T., Lega, L. & Feitelson, M. A. ( 1991b; ). HBxAg in liver from carrier patients with chronic hepatitis and cirrhosis. Hepatology 14, 29-37.[CrossRef]
    [Google Scholar]
  73. Wang, X. W., Gibson, M. K., Vermeulen, W., Yeh, H., Forrester, K., Sturzbecher, H. W., Hoeijmakers, J. H. J. & Harris, C. C. ( 1995; ). Abrogation of p53-induced apoptosis by the HBV X gene. Cancer Research 55, 6012-6016.
    [Google Scholar]
  74. Wang, C.-Y., Mayo, M. W. & Baldwin, A. S.Jr ( 1996; ). TNF-α and cancer therapy-induced apoptosis: potentiation by inhibition of NF-κB. Science 274, 784-786.[CrossRef]
    [Google Scholar]
  75. Watanabe-Fukunage, R., Brannan, C. I., Itoh, N., Yonehara, S., Copeland, N. G., Jenkins, N. A. & Nagata, S. ( 1992; ). The cDNA structure, expression, chromosomal assignment of the mouse Fas antigen. Journal of Immunology 148, 1274-1279.
    [Google Scholar]
  76. Weil, R., Sirma, H., Giannini, C., Kremsdorf, D., Bessia, C., Dargemont, C., Brechot, C. & Israel, A. ( 1999; ). Direct association and nuclear import of the hepatitis B virus X protein with the NF-κB inhibitor IκBα. Molecular and Cellular Biology 19, 6345-6354.
    [Google Scholar]
  77. Wollersheim, M., Debelka, U. & Hofschneider, P. H. ( 1988; ). A transactivating function encoded in the hepatitis B virus X gene is conserved in the integrated state. Oncogene 3, 545-552.
    [Google Scholar]
  78. Zahm, P., Hofschneider, P. H. & Koshy, R. ( 1988; ). The HBV X-ORF encodes a transactivator: a potential factor in viral hepatocarcinogenesis. Oncogene 3, 169-177.
    [Google Scholar]
  79. Zong, W. X., Bash, J. & Gelinas, C. ( 1998; ). Rel blocks both anti-Fas- and TNF alpha-induced apoptosis and an intact Rel transactivation domain is essential for this effect. Cell Death & Differentiation 5, 963-972.[CrossRef]
    [Google Scholar]
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