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. 1996 Jul;70(7):4438–4443. doi: 10.1128/jvi.70.7.4438-4443.1996

Hepatitis C virus core protein cooperates with ras and transforms primary rat embryo fibroblasts to tumorigenic phenotype.

R B Ray 1, L M Lagging 1, K Meyer 1, R Ray 1
PMCID: PMC190377  PMID: 8676467

Abstract

We have previously demonstrated that hepatitis C virus (HCV) core protein regulates cellular protooncogenes at the transcriptional level; this observation implicates core protein in the alteration of normal hepatocyte growth. In the present study, the transforming potential of the HCV core gene was investigated by using primary rat embryo fibroblast (REF) cells which were transfected with or without cooperative oncogenes. Integration of the HCV core gene resulted in expression of the viral protein in REF stable transformants. REF cells cotransfected with HCV core and H-ras genes became transformed and exhibited rapid proliferation, anchor-independent growth, and tumor formation in athymic nude mice. Results from these studies suggest that the core protein plays an important role in the regulation of HCV-infected cell growth and in the transformation to tumorigenic phenotype. These observations suggest a possible mechanism for this viral protein in the pathogenesis of hepatocellular carcinoma in HCV-infected humans.

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Selected References

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  1. Alter H. J., Purcell R. H., Shih J. W., Melpolder J. C., Houghton M., Choo Q. L., Kuo G. Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. N Engl J Med. 1989 Nov 30;321(22):1494–1500. doi: 10.1056/NEJM198911303212202. [DOI] [PubMed] [Google Scholar]
  2. Birrer M. J., Segal S., DeGreve J. S., Kaye F., Sausville E. A., Minna J. D. L-myc cooperates with ras to transform primary rat embryo fibroblasts. Mol Cell Biol. 1988 Jun;8(6):2668–2673. doi: 10.1128/mcb.8.6.2668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bishop J. M. The molecular genetics of cancer. Science. 1987 Jan 16;235(4786):305–311. doi: 10.1126/science.3541204. [DOI] [PubMed] [Google Scholar]
  4. Brough D. E., Hofmann T. J., Ellwood K. B., Townley R. A., Cole M. D. An essential domain of the c-myc protein interacts with a nuclear factor that is also required for E1A-mediated transformation. Mol Cell Biol. 1995 Mar;15(3):1536–1544. doi: 10.1128/mcb.15.3.1536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bruix J., Barrera J. M., Calvet X., Ercilla G., Costa J., Sanchez-Tapias J. M., Ventura M., Vall M., Bruguera M., Bru C. Prevalence of antibodies to hepatitis C virus in Spanish patients with hepatocellular carcinoma and hepatic cirrhosis. Lancet. 1989 Oct 28;2(8670):1004–1006. doi: 10.1016/s0140-6736(89)91015-5. [DOI] [PubMed] [Google Scholar]
  6. Bukh J., Purcell R. H., Miller R. H. Sequence analysis of the core gene of 14 hepatitis C virus genotypes. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8239–8243. doi: 10.1073/pnas.91.17.8239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cerni C., Binétruy B., Schiller J. T., Lowy D. R., Meneguzzi G., Cuzin F. Successive steps in the process of immortalization identified by transfer of separate bovine papillomavirus genes into rat fibroblasts. Proc Natl Acad Sci U S A. 1989 May;86(9):3266–3270. doi: 10.1073/pnas.86.9.3266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cerny A., Chisari F. V. Immunological aspects of HCV infection. Intervirology. 1994;37(2):119–125. doi: 10.1159/000150366. [DOI] [PubMed] [Google Scholar]
  9. Chien D. Y., Choo Q. L., Ralston R., Spaete R., Tong M., Houghton M., Kuo G. Persistence of HCV despite antibodies to both putative envelope glycoproteins. Lancet. 1993 Oct 9;342(8876):933–933. doi: 10.1016/0140-6736(93)91983-s. [DOI] [PubMed] [Google Scholar]
  10. Choo Q. L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science. 1989 Apr 21;244(4902):359–362. doi: 10.1126/science.2523562. [DOI] [PubMed] [Google Scholar]
  11. Choo Q. L., Richman K. H., Han J. H., Berger K., Lee C., Dong C., Gallegos C., Coit D., Medina-Selby R., Barr P. J. Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2451–2455. doi: 10.1073/pnas.88.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Colombo M., Kuo G., Choo Q. L., Donato M. F., Del Ninno E., Tommasini M. A., Dioguardi N., Houghton M. Prevalence of antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma. Lancet. 1989 Oct 28;2(8670):1006–1008. doi: 10.1016/s0140-6736(89)91016-7. [DOI] [PubMed] [Google Scholar]
  13. Crook T., Storey A., Almond N., Osborn K., Crawford L. Human papillomavirus type 16 cooperates with activated ras and fos oncogenes in the hormone-dependent transformation of primary mouse cells. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8820–8824. doi: 10.1073/pnas.85.23.8820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Farci P., Alter H. J., Wong D. C., Miller R. H., Govindarajan S., Engle R., Shapiro M., Purcell R. H. Prevention of hepatitis C virus infection in chimpanzees after antibody-mediated in vitro neutralization. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7792–7796. doi: 10.1073/pnas.91.16.7792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hamburger A. W., Salmon S. E. Development of a bioassay for human myeloma colony-forming cells. Prog Clin Biol Res. 1980;48:23–41. [PubMed] [Google Scholar]
  16. Hawley-Nelson P., Vousden K. H., Hubbert N. L., Lowy D. R., Schiller J. T. HPV16 E6 and E7 proteins cooperate to immortalize human foreskin keratinocytes. EMBO J. 1989 Dec 1;8(12):3905–3910. doi: 10.1002/j.1460-2075.1989.tb08570.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jewers R. J., Hildebrandt P., Ludlow J. W., Kell B., McCance D. J. Regions of human papillomavirus type 16 E7 oncoprotein required for immortalization of human keratinocytes. J Virol. 1992 Mar;66(3):1329–1335. doi: 10.1128/jvi.66.3.1329-1335.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kelekar A., Cole M. D. Immortalization by c-myc, H-ras, and Ela oncogenes induces differential cellular gene expression and growth factor responses. Mol Cell Biol. 1987 Nov;7(11):3899–3907. doi: 10.1128/mcb.7.11.3899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kew M. C., Houghton M., Choo Q. L., Kuo G. Hepatitis C virus antibodies in southern African blacks with hepatocellular carcinoma. Lancet. 1990 Apr 14;335(8694):873–874. doi: 10.1016/0140-6736(90)90474-j. [DOI] [PubMed] [Google Scholar]
  20. Kew M. C. Tumours of the liver. Scand J Gastroenterol Suppl. 1992;192:39–42. doi: 10.3109/00365529209095977. [DOI] [PubMed] [Google Scholar]
  21. Khanna A., Ray R. Hepatitis C virus core protein: synthesis, affinity purification and immunoreactivity with infected human sera. Gene. 1995 Feb 14;153(2):185–189. doi: 10.1016/0378-1119(94)00782-n. [DOI] [PubMed] [Google Scholar]
  22. Lagging L. M., Meyer K., Hoft D., Houghton M., Belshe R. B., Ray R. Immune responses to plasmid DNA encoding the hepatitis C virus core protein. J Virol. 1995 Sep;69(9):5859–5863. doi: 10.1128/jvi.69.9.5859-5863.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Land H., Chen A. C., Morgenstern J. P., Parada L. F., Weinberg R. A. Behavior of myc and ras oncogenes in transformation of rat embryo fibroblasts. Mol Cell Biol. 1986 Jun;6(6):1917–1925. doi: 10.1128/mcb.6.6.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Land H., Parada L. F., Weinberg R. A. Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature. 1983 Aug 18;304(5927):596–602. doi: 10.1038/304596a0. [DOI] [PubMed] [Google Scholar]
  25. Lanford R. E., Chavez D., Chisari F. V., Sureau C. Lack of detection of negative-strand hepatitis C virus RNA in peripheral blood mononuclear cells and other extrahepatic tissues by the highly strand-specific rTth reverse transcriptase PCR. J Virol. 1995 Dec;69(12):8079–8083. doi: 10.1128/jvi.69.12.8079-8083.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Liu Z., Ghai J., Ostrow R. S., McGlennen R. C., Faras A. J. The E6 gene of human papillomavirus type 16 is sufficient for transformation of baby rat kidney cells in cotransfection with activated Ha-ras. Virology. 1994 Jun;201(2):388–396. doi: 10.1006/viro.1994.1306. [DOI] [PubMed] [Google Scholar]
  27. Lo S. Y., Masiarz F., Hwang S. B., Lai M. M., Ou J. H. Differential subcellular localization of hepatitis C virus core gene products. Virology. 1995 Nov 10;213(2):455–461. doi: 10.1006/viro.1995.0018. [DOI] [PubMed] [Google Scholar]
  28. Matlashewski G., Schneider J., Banks L., Jones N., Murray A., Crawford L. Human papillomavirus type 16 DNA cooperates with activated ras in transforming primary cells. EMBO J. 1987 Jun;6(6):1741–1746. doi: 10.1002/j.1460-2075.1987.tb02426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Münger K., Phelps W. C., Bubb V., Howley P. M., Schlegel R. The E6 and E7 genes of the human papillomavirus type 16 together are necessary and sufficient for transformation of primary human keratinocytes. J Virol. 1989 Oct;63(10):4417–4421. doi: 10.1128/jvi.63.10.4417-4421.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Purcell R. H. Hepatitis viruses: changing patterns of human disease. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2401–2406. doi: 10.1073/pnas.91.7.2401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ray R. B. Induction of cell death in murine fibroblasts by a c-myc promoter binding protein. Cell Growth Differ. 1995 Sep;6(9):1089–1096. [PubMed] [Google Scholar]
  32. Ray R. B., Lagging L. M., Meyer K., Steele R., Ray R. Transcriptional regulation of cellular and viral promoters by the hepatitis C virus core protein. Virus Res. 1995 Aug;37(3):209–220. doi: 10.1016/0168-1702(95)00034-n. [DOI] [PubMed] [Google Scholar]
  33. Ray R. B., Steele R., Seftor E., Hendrix M. Human breast carcinoma cells transfected with the gene encoding a c-myc promoter-binding protein (MBP-1) inhibits tumors in nude mice. Cancer Res. 1995 Sep 1;55(17):3747–3751. [PubMed] [Google Scholar]
  34. Ray R., Khanna A., Lagging L. M., Meyer K., Choo Q. L., Ralston R., Houghton M., Becherer P. R. Peptide immunogen mimicry of putative E1 glycoprotein-specific epitopes in hepatitis C virus. J Virol. 1994 Jul;68(7):4420–4426. doi: 10.1128/jvi.68.7.4420-4426.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ray R., Miller D. M. Cloning and characterization of a human c-myc promoter-binding protein. Mol Cell Biol. 1991 Apr;11(4):2154–2161. doi: 10.1128/mcb.11.4.2154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ray R., Thomas S., Miller D. M. Mouse fibroblasts transformed with the human c-myc gene express a high level of mRNA but a low level of c-myc protein and are non-tumorigenic in nude mice. Oncogene. 1989 May;4(5):593–600. [PubMed] [Google Scholar]
  37. Rogler C. E., Chisari F. V. Cellular and molecular mechanisms of hepatocarcinogenesis. Semin Liver Dis. 1992 Aug;12(3):265–278. doi: 10.1055/s-2007-1007398. [DOI] [PubMed] [Google Scholar]
  38. Rovinski B., Benchimol S. Immortalization of rat embryo fibroblasts by the cellular p53 oncogene. Oncogene. 1988 May;2(5):445–452. [PubMed] [Google Scholar]
  39. Ruley H. E. Adenovirus early region 1A enables viral and cellular transforming genes to transform primary cells in culture. Nature. 1983 Aug 18;304(5927):602–606. doi: 10.1038/304602a0. [DOI] [PubMed] [Google Scholar]
  40. Saito I., Miyamura T., Ohbayashi A., Harada H., Katayama T., Kikuchi S., Watanabe Y., Koi S., Onji M., Ohta Y. Hepatitis C virus infection is associated with the development of hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6547–6549. doi: 10.1073/pnas.87.17.6547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sakamuro D., Furukawa T., Takegami T. Hepatitis C virus nonstructural protein NS3 transforms NIH 3T3 cells. J Virol. 1995 Jun;69(6):3893–3896. doi: 10.1128/jvi.69.6.3893-3896.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Santolini E., Migliaccio G., La Monica N. Biosynthesis and biochemical properties of the hepatitis C virus core protein. J Virol. 1994 Jun;68(6):3631–3641. doi: 10.1128/jvi.68.6.3631-3641.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Selby M. J., Choo Q. L., Berger K., Kuo G., Glazer E., Eckart M., Lee C., Chien D., Kuo C., Houghton M. Expression, identification and subcellular localization of the proteins encoded by the hepatitis C viral genome. J Gen Virol. 1993 Jun;74(Pt 6):1103–1113. doi: 10.1099/0022-1317-74-6-1103. [DOI] [PubMed] [Google Scholar]
  44. Shenk T., Flint J. Transcriptional and transforming activities of the adenovirus E1A proteins. Adv Cancer Res. 1991;57:47–85. doi: 10.1016/s0065-230x(08)60995-1. [DOI] [PubMed] [Google Scholar]
  45. Shih C. M., Chen C. M., Chen S. Y., Lee Y. H. Modulation of the trans-suppression activity of hepatitis C virus core protein by phosphorylation. J Virol. 1995 Feb;69(2):1160–1171. doi: 10.1128/jvi.69.2.1160-1171.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Shih C. M., Lo S. J., Miyamura T., Chen S. Y., Lee Y. H. Suppression of hepatitis B virus expression and replication by hepatitis C virus core protein in HuH-7 cells. J Virol. 1993 Oct;67(10):5823–5832. doi: 10.1128/jvi.67.10.5823-5832.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Smotkin D., Wettstein F. O. The major human papillomavirus protein in cervical cancers is a cytoplasmic phosphoprotein. J Virol. 1987 May;61(5):1686–1689. doi: 10.1128/jvi.61.5.1686-1689.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Suzuki R., Matsuura Y., Suzuki T., Ando A., Chiba J., Harada S., Saito I., Miyamura T. Nuclear localization of the truncated hepatitis C virus core protein with its hydrophobic C terminus deleted. J Gen Virol. 1995 Jan;76(Pt 1):53–61. doi: 10.1099/0022-1317-76-1-53. [DOI] [PubMed] [Google Scholar]
  49. Tappero G., Natoli G., Anfossi G., Rosina F., Negro F., Smedile A., Bonino F., Angeli A., Purcell R. H., Rizzetto M. Expression of the c-myc protooncogene product in cells infected with the hepatitis delta virus. Hepatology. 1994 Nov;20(5):1109–1114. [PubMed] [Google Scholar]
  50. Teramoto T., Satonaka K., Kitazawa S., Fujimori T., Hayashi K., Maeda S. p53 gene abnormalities are closely related to hepatoviral infections and occur at a late stage of hepatocarcinogenesis. Cancer Res. 1994 Jan 1;54(1):231–235. [PubMed] [Google Scholar]
  51. Werness B. A., Levine A. J., Howley P. M. Association of human papillomavirus types 16 and 18 E6 proteins with p53. Science. 1990 Apr 6;248(4951):76–79. doi: 10.1126/science.2157286. [DOI] [PubMed] [Google Scholar]

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