1887

Abstract

Infection by hepatitis B virus (HBV) and dietary exposure to aflatoxin B (AFB) are the main risk factors for the development of chronic liver disease and hepatocellular carcinoma (HCC). How these factors cooperate is still largely unknown. AFB activation leads to DNA adduction and mutagenesis, with a specific mutation at codon 249 in (p.R249S). So far, only limited studies have addressed the effects of AFB on HBV replication. We have analysed the effects of both risk factors on p53 induction during HBV infection in HepaRG, a cell line with hepatocyte-like morphology that metabolizes AFB and supports HBV infection. Exposure to AFB up to 5 µM induced a downregulation of HBV replication after 48 h, as measured by a decrease in viral antigens in the culture medium (HBsAg, HBeAg and large envelope protein) and in intracellular levels of HBV transcripts, DNA and HBsAg. Conversely, HBV infection did not significantly modify AFB-DNA adduct formation or repair as assessed by immunodot-blot assay, and the induction of p53 in response to AFB was similar in infected and non-infected HepaRG cells. Overall, our results suggest that AFB exposure decreases HBV replication, whereas DNA damage by AFB and subsequent p53 induction is not affected by the presence of the virus. Thus, in HepaRG cell line, AFB and HBV do not cooperate to increase DNA damage by AFB. Further studies on the effects of both factors in a context of chronicity are needed to better understand synergistic effects.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.032482-0
2012-03-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/93/3/640.html?itemId=/content/journal/jgv/10.1099/vir.0.032482-0&mimeType=html&fmt=ahah

References

  1. Aguilar F., Hussain S. P., Cerutti P. 1993; Aflatoxin B1 induces the transversion of G→T in codon 249 of the p53 tumor suppressor gene in human hepatocytes. Proc Natl Acad Sci U S A 90:8586–8590 [View Article][PubMed]
    [Google Scholar]
  2. Andrieux L. O., Fautrel A., Bessard A., Guillouzo A., Baffet G., Langouët S. 2007; GATA-1 is essential in EGF-mediated induction of nucleotide excision repair activity and ERCC1 expression through ERK2 in human hepatoma cells. Cancer Res 67:2114–2123 [View Article][PubMed]
    [Google Scholar]
  3. Aninat C., Piton A., Glaise D., Le Charpentier T., Langouët S., Morel F., Guguen-Guillouzo C., Guillouzo A. 2006; Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells. Drug Metab Dispos 34:75–83 [View Article][PubMed]
    [Google Scholar]
  4. Barraud L., Guerret S., Chevallier M., Borel C., Jamard C., Trepo C., Wild C. P., Cova L. 1999; Enhanced duck hepatitis B virus gene expression following aflatoxin B1 exposure. Hepatology 29:1317–1323 [View Article][PubMed]
    [Google Scholar]
  5. Barraud L., Douki T., Guerret S., Chevallier M., Jamard C., Trepo C., Wild C. P., Cadet J., Cova L. 2001; The role of duck hepatitis B virus and aflatoxin B1 in the induction of oxidative stress in the liver. Cancer Detect Prev 25:192–201[PubMed]
    [Google Scholar]
  6. Besaratinia A., Kim S. I., Hainaut P., Pfeifer G. P. 2009; In vitro recapitulating of TP53 mutagenesis in hepatocellular carcinoma associated with dietary aflatoxin B1 exposure. Gastroenterology 137:1127–1137, 1137.e5 [View Article][PubMed]
    [Google Scholar]
  7. Bosch F. X., Ribes J., Borràs J. 1999; Epidemiology of primary liver cancer. Semin Liver Dis 19:271–285 [View Article][PubMed]
    [Google Scholar]
  8. Bressac B., Kew M., Wands J., Ozturk M. 1991; Selective G to T mutations of p53 gene in hepatocellular carcinoma from southern Africa. Nature 350:429–431 [View Article][PubMed]
    [Google Scholar]
  9. Chemin I., Ohgaki H., Chisari F. V., Wild C. P. 1999; Altered expression of hepatic carcinogen metabolizing enzymes with liver injury in HBV transgenic mouse lineages expressing various amounts of hepatitis B surface antigen. Liver 19:81–87 [View Article][PubMed]
    [Google Scholar]
  10. Ferlay J., Shin H. R., Bray F., Forman D., Mathers C., Parkin D. M. 2010 GLOBOCAN 2008, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10 [Internet]: Lyon, France: International Agency for Research on Cancer. Available from http://globocan.iarc.fr
  11. Ganem D., Prince A. M. 2004; Hepatitis B virus infection–natural history and clinical consequences. N Engl J Med 350:1118–1129 [View Article][PubMed]
    [Google Scholar]
  12. Gouas D., Shi H., Hainaut P. 2009; The aflatoxin-induced TP53 mutation at codon 249 (R249S): biomarker of exposure, early detection and target for therapy. Cancer Lett 286:29–37 [View Article][PubMed]
    [Google Scholar]
  13. Gripon P., Rumin S., Urban S., Le Seyec J., Glaise D., Cannie I., Guyomard C., Lucas J., Trepo C., Guguen-Guillouzo C. 2002; Infection of a human hepatoma cell line by hepatitis B virus. Proc Natl Acad Sci U S A 99:15655–15660 [View Article][PubMed]
    [Google Scholar]
  14. Hantz O., Parent R., Durantel D., Gripon P., Guguen-Guillouzo C., Zoulim F. 2009; Persistence of the hepatitis B virus covalently closed circular DNA in HepaRG human hepatocyte-like cells. J Gen Virol 90:127–135 [View Article][PubMed]
    [Google Scholar]
  15. Hollstein M., Hainaut P. 2010; Massively regulated genes: the example of TP53. J Pathol 220:164–173[PubMed]
    [Google Scholar]
  16. Hsu I. C., Metcalf R. A., Sun T., Welsh J. A., Wang N. J., Harris C. C. 1991; Mutational hotspot in the p53 gene in human hepatocellular carcinomas. Nature 350:427–428 [View Article][PubMed]
    [Google Scholar]
  17. Jennen D. G., Magkoufopoulou C., Ketelslegers H. B., van Herwijnen M. H., Kleinjans J. C., van Delft J. H. 2010; Comparison of HepG2 and HepaRG by whole-genome gene expression analysis for the purpose of chemical hazard identification. Toxicol Sci 115:66–79 [View Article][PubMed]
    [Google Scholar]
  18. Jia L., Wang X. W., Harris C. C. 1999; Hepatitis B virus X protein inhibits nucleotide excision repair. Int J Cancer 80:875–879 [View Article][PubMed]
    [Google Scholar]
  19. Kew M. C. 2003; Synergistic interaction between aflatoxin B1 and hepatitis B virus in hepatocarcinogenesis. Liver Int 23:405–409 [View Article][PubMed]
    [Google Scholar]
  20. Le Guillou D. B., Duclos-Vallée J. C., Eberle F., Capel F., Petit M. A. 2000; Evaluation of an enzyme-linked immunosorbent assay for detection and quantification of hepatitis B virus preS1 envelope antigen in serum samples: comparison with two commercial assays for monitoring hepatitis B virus DNA. J Viral Hepat 7:387–392 [View Article][PubMed]
    [Google Scholar]
  21. Lleonart M. E., Kirk G. D., Villar S., Lesi O. A., Dasgupta A., Goedert J. J., Mendy M., Hollstein M. C., Montesano R. other authors 2005; Quantitative analysis of plasma TP53 249Ser-mutated DNA by electrospray ionization mass spectrometry. Cancer Epidemiol Biomarkers Prev 14:2956–2962 [View Article][PubMed]
    [Google Scholar]
  22. Lucifora J., Arzberger S., Durantel D., Belloni L., Strubin M., Levrero M., Zoulim F., Hantz O., Protzer U. 2011; Hepatitis B virus X protein is essential to initiate and maintain virus replication after infection. J Hepatol 55:996–1003 [View Article][PubMed]
    [Google Scholar]
  23. Lunn R. M., Zhang Y. J., Wang L. Y., Chen C. J., Lee P. H., Lee C. S., Tsai W. Y., Santella R. M. 1997; p53 mutations, chronic hepatitis B virus infection, and aflatoxin exposure in hepatocellular carcinoma in Taiwan. Cancer Res 57:3471–3477[PubMed]
    [Google Scholar]
  24. McGlynn K. A., London W. T. 2005; Epidemiology and natural history of hepatocellular carcinoma. Best Pract Res Clin Gastroenterol 19:3–23 [View Article][PubMed]
    [Google Scholar]
  25. Ozturk M. 1991; p53 mutation in hepatocellular carcinoma after aflatoxin exposure. Lancet 338:1356–1359 [View Article][PubMed]
    [Google Scholar]
  26. Qian G. S., Ross R. K., Yu M. C., Yuan J. M., Gao Y. T., Henderson B. E., Wogan G. N., Groopman J. D. 1994; A follow-up study of urinary markers of aflatoxin exposure and liver cancer risk in Shanghai, People’s Republic of China. Cancer Epidemiol Biomarkers Prev 3:3–10[PubMed]
    [Google Scholar]
  27. Ross R. K., Yu M. C., Henderson B. E., Yuan J.-M., Qian G.-S., Tu J.-T., Gao Y.-T., Wogan G. N., Groopman J. D. 1992; Urinary aflatoxin biomarkers and risk of hepatocellular carcinoma. Lancet 339:943–946 [View Article][PubMed]
    [Google Scholar]
  28. Sarasin A. R., Smith C. A., Hanawalt P. C. 1977; Repair of DNA in human cells after treatment with activated aflatoxin B1. Cancer Res 37:1786–1793[PubMed]
    [Google Scholar]
  29. Sells M. A., Chen M. L., Acs G. 1987; Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci U S A 84:1005–1009 [View Article][PubMed]
    [Google Scholar]
  30. Smela M. E., Hamm M. L., Henderson P. T., Harris C. M., Harris T. M., Essigmann J. M. 2002; The aflatoxin B1 formamidopyrimidine adduct plays a major role in causing the types of mutations observed in human hepatocellular carcinoma. Proc Natl Acad Sci U S A 99:6655–6660 [View Article][PubMed]
    [Google Scholar]
  31. Sylla A., Diallo M. S., Castegnaro J., Wild C. P. 1999; Interactions between hepatitis B virus infection and exposure to aflatoxins in the development of hepatocellular carcinoma: a molecular epidemiological approach. Mutat Res 428:187–196 [View Article][PubMed]
    [Google Scholar]
  32. Untergasser A., Zedler U., Langenkamp A., Hösel M., Quasdorff M., Esser K., Dienes H. P., Tappertzhofen B., Kolanus W., Protzer U. 2006; Dendritic cells take up viral antigens but do not support the early steps of hepatitis B virus infection. Hepatology 43:539–547 [View Article][PubMed]
    [Google Scholar]
  33. Wang L. Y., Hatch M., Chen C. J., Levin B., You S. L., Lu S. N., Wu M. H., Wu W. P., Wang L. W. other authors 1996; Aflatoxin exposure and risk of hepatocellular carcinoma in Taiwan. Int J Cancer 67:620–625 [View Article][PubMed]
    [Google Scholar]
  34. Waters R., Jones C. J., Martin E. A., Yang A. L., Jones N. J. 1992; The repair of large DNA adducts in mammalian cells. Mutat Res 273:145–155[PubMed] [CrossRef]
    [Google Scholar]
  35. WHO 2002 Evaluation of carcinogenic risks to humans. Some traditional herbal medicines, some mycotoxins, naphthalene and styrene vol. 82 pp. 301–366 Lyon, France: IARC Press;
    [Google Scholar]
  36. Williams J. H., Phillips T. D., Jolly P. E., Stiles J. K., Jolly C. M., Aggarwal D. 2004; Human aflatoxicosis in developing countries: a review of toxicology, exposure, potential health consequences, and interventions. Am J Clin Nutr 80:1106–1122[PubMed]
    [Google Scholar]
  37. Zhang Y. J., Chen C. J., Lee C. S., Haghighi B., Yang G. Y., Wang L. W., Feitelson M., Santella R. 1991; Aflatoxin B1-DNA adducts and hepatitis B virus antigens in hepatocellular carcinoma and non-tumorous liver tissue. Carcinogenesis 12:2247–2252 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.032482-0
Loading
/content/journal/jgv/10.1099/vir.0.032482-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error