1887

Abstract

Hepatitis B virus X protein (HBx) is involved in the development of hepatocellular carcinoma (HCC). The HBx sequence is a preferential site of integration into the human genome, leading to the formation of C-terminal-truncated HBx proteins (Ct-HBx). We previously reported that Ct-HBx proteins were able to potentiate cell transformation . Our present goal was to compare the ability of Ct-HBx and full-length HBx (FL-HBx) proteins to develop or enhance HCC in transgenic mice. In the absence of treatment, neither Ct-HBx- nor FL-HBx-transgenic mice developed HCC. In young mice treated with diethylnitrosamine (DEN) at 8 months of age, a significantly higher incidence and number of liver lesions were observed in Ct-HBx mice than in FL-HBx and control mice. The earlier development of tumours in Ct-HBx-transgenic mice was associated with increased liver inflammation. At 10 months, macroscopic and microscopic analyses showed that, statistically, FL-HBx mice developed more liver lesions with a larger surface area than control mice. Furthermore, during DEN-induced initiation of HCC, Ct-HBx- and FL-HBx-transgenic mice showed higher expression of IL-6, TNF-α and IL-1β transcripts, activation of STAT3, ERK and JNK proteins and an increase in cell apoptosis. In conclusion, in DEN-treated transgenic mice, the expression of Ct-HBx protein causes a more rapid onset of HCC than does FL-HBx protein. HBV genome integration leading to the expression of a truncated form of HBx protein may therefore facilitate HCC development in chronically infected patients.

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2015-03-01
2020-09-25
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References

  1. Assrir N., Soussan P., Kremsdorf D., Rossignol J. M. 2010; Role of the hepatitis B virus proteins in pro- and anti-apoptotic processes. Front Biosci 15:12–24 [CrossRef][PubMed]
    [Google Scholar]
  2. Bergametti F., Sitterlin D., Transy C. 2002; Turnover of hepatitis B virus X protein is regulated by damaged DNA-binding complex. J Virol 76:6495–6501 [CrossRef][PubMed]
    [Google Scholar]
  3. Bock C. T., Toan N. L., Koeberlein B., Song L. H., Chin R., Zentgraf H., Kandolf R., Torresi J. 2008; Subcellular mislocalization of mutant hepatitis B X proteins contributes to modulation of STAT/SOCS signaling in hepatocellular carcinoma. Intervirology 51:432–443 [CrossRef][PubMed]
    [Google Scholar]
  4. Brandon-Warner E., Walling T. L., Schrum L. W., McKillop I. H. 2012; Chronic ethanol feeding accelerates hepatocellular carcinoma progression in a sex-dependent manner in a mouse model of hepatocarcinogenesis. Alcohol Clin Exp Res 36:641–653 [CrossRef][PubMed]
    [Google Scholar]
  5. Brechot C., Kremsdorf D., Soussan P., Pineau P., Dejean A., Paterlini-Brechot P., Tiollais P. 2010; Hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC): molecular mechanisms and novel paradigms. Pathol Biol (Paris) 58:278–287 [CrossRef][PubMed]
    [Google Scholar]
  6. Fu X., Tan D., Hou Z., Hu Z., Liu G., Ouyang Y., Liu F. 2012; The effect of miR-338-3p on HBx deletion-mutant (HBx-d382) mediated liver-cell proliferation through CyclinD1 regulation. PLoS ONE 7:e43204 [CrossRef][PubMed]
    [Google Scholar]
  7. He G., Yu G. Y., Temkin V., Ogata H., Kuntzen C., Sakurai T., Sieghart W., Peck-Radosavljevic M., Leffert H. L., Karin M. 2010; Hepatocyte IKK β/NF-κB inhibits tumor promotion and progression by preventing oxidative stress-driven STAT3 activation. Cancer Cell 17:286–297 [CrossRef][PubMed]
    [Google Scholar]
  8. Huo T. I., Wang X. W., Forgues M., Wu C. G., Spillare E. A., Giannini C., Brechot C., Harris C. C. 2001; Hepatitis B virus X mutants derived from human hepatocellular carcinoma retain the ability to abrogate p53-induced apoptosis. Oncogene 20:3620–3628 [CrossRef][PubMed]
    [Google Scholar]
  9. Iavarone M., Trabut J. B., Delpuech O., Carnot F., Colombo M., Kremsdorf D., Bréchot C., Thiers V. 2003; Characterisation of hepatitis B virus X protein mutants in tumour and non-tumour liver cells using laser capture microdissection. J Hepatol 39:253–261 [CrossRef][PubMed]
    [Google Scholar]
  10. Jiang W., Wang X. W., Unger T., Forgues M., Kim J. W., Hussain S. P., Bowman E., Spillare E. A., Lipsky M. M. other authors 2010; Cooperation of tumor-derived HBx mutants and p53-249(ser) mutant in regulating cell proliferation, anchorage-independent growth and aneuploidy in a telomerase-immortalized normal human hepatocyte-derived cell line. Int J Cancer 127:1011–1020 [CrossRef][PubMed]
    [Google Scholar]
  11. Jiang Z., Jhunjhunwala S., Liu J., Haverty P. M., Kennemer M. I., Guan Y., Lee W., Carnevali P., Stinson J. other authors 2012; The effects of hepatitis B virus integration into the genomes of hepatocellular carcinoma patients. Genome Res 22:593–601 [CrossRef][PubMed]
    [Google Scholar]
  12. Jung S. Y., Kim Y. J. 2013; C-terminal region of HBx is crucial for mitochondrial DNA damage. Cancer Lett 331:76–83 [CrossRef][PubMed]
    [Google Scholar]
  13. Kew M. C. 2011; Hepatitis B virus x protein in the pathogenesis of hepatitis B virus-induced hepatocellular carcinoma. J Gastroenterol Hepatol 26:Suppl. 1144–152 [CrossRef][PubMed]
    [Google Scholar]
  14. Kremsdorf D., Soussan P., Paterlini-Brechot P., Brechot C. 2006; Hepatitis B virus-related hepatocellular carcinoma: paradigms for viral-related human carcinogenesis. Oncogene 25:3823–3833 [CrossRef][PubMed]
    [Google Scholar]
  15. Kuang S. Y., Jackson P. E., Wang J. B., Lu P. X., Muñoz A., Qian G. S., Kensler T. W., Groopman J. D. 2004; Specific mutations of hepatitis B virus in plasma predict liver cancer development. Proc Natl Acad Sci U S A 101:3575–3580 [CrossRef][PubMed]
    [Google Scholar]
  16. Lee J. S., Chu I. S., Mikaelyan A., Calvisi D. F., Heo J., Reddy J. K., Thorgeirsson S. S. 2004; Application of comparative functional genomics to identify best-fit mouse models to study human cancer. Nat Genet 36:1306–1311 [CrossRef][PubMed]
    [Google Scholar]
  17. Lee J. H., Han K. H., Lee J. M., Park J. H., Kim H. S. 2011; Impact of hepatitis B virus (HBV) x gene mutations on hepatocellular carcinoma development in chronic HBV infection. Clin Vaccine Immunol 18:914–921 [CrossRef][PubMed]
    [Google Scholar]
  18. Li M., Zhu M., Li W., Lu Y., Xie X., Wu Y., Zheng S. 2013; Alpha-fetoprotein receptor as an early indicator of HBx-driven hepatocarcinogenesis and its applications in tracing cancer cell metastasis. Cancer Lett 330:170–180 [CrossRef][PubMed]
    [Google Scholar]
  19. Lin-Marq N., Bontron S., Leupin O., Strubin M. 2001; Hepatitis B virus X protein interferes with cell viability through interaction with the p127-kDa UV-damaged DNA-binding protein. Virology 287:266–274 [CrossRef][PubMed]
    [Google Scholar]
  20. Liu X., Wang L., Zhang S., Lin J., Zhang S., Feitelson M. A., Gao H., Zhu M. 2008a; Mutations in the C-terminus of the X protein of hepatitis B virus regulate Wnt-5a expression in hepatoma Huh7 cells: cDNA microarray and proteomic analyses. Carcinogenesis 29:1207–1214 [CrossRef][PubMed]
    [Google Scholar]
  21. Liu X., Zhang S., Lin J., Zhang S., Feitelson M. A., Gao H., Zhu M. 2008b; Hepatitis B virus X protein mutants exhibit distinct biological activities in hepatoma Huh7 cells. Biochem Biophys Res Commun 373:643–647 [CrossRef][PubMed]
    [Google Scholar]
  22. Ma N. F., Lau S. H., Hu L., Xie D., Wu J., Yang J., Wang Y., Wu M. C., Fung J. other authors 2008; COOH-terminal truncated HBV X protein plays key role in hepatocarcinogenesis. Clin Cancer Res 14:5061–5068 [CrossRef][PubMed]
    [Google Scholar]
  23. Madden C. R., Finegold M. J., Slagle B. L. 2001; Hepatitis B virus X protein acts as a tumor promoter in development of diethylnitrosamine-induced preneoplastic lesions. J Virol 75:3851–3858 [CrossRef][PubMed]
    [Google Scholar]
  24. Maeda S., Kamata H., Luo J. L., Leffert H., Karin M. 2005; IKKβ couples hepatocyte death to cytokine-driven compensatory proliferation that promotes chemical hepatocarcinogenesis. Cell 121:977–990 [CrossRef][PubMed]
    [Google Scholar]
  25. Malmassari S. L., Deng Q., Fontaine H., Houitte D., Rimlinger F., Thiers V., Maillere B., Pol S., Michel M. L. 2007; Impact of hepatitis B virus basic core promoter mutations on T cell response to an immunodominant HBx-derived epitope. Hepatology 45:1199–1209 [CrossRef][PubMed]
    [Google Scholar]
  26. Min L., He B., Hui L. 2011; Mitogen-activated protein kinases in hepatocellular carcinoma development. Semin Cancer Biol 21:10–20 [CrossRef][PubMed]
    [Google Scholar]
  27. Naugler W. E., Sakurai T., Kim S., Maeda S., Kim K., Elsharkawy A. M., Karin M. 2007; Gender disparity in liver cancer due to sex differences in MyD88-dependent IL-6 production. Science 317:121–124 [CrossRef][PubMed]
    [Google Scholar]
  28. Nijhara R., Jana S. S., Goswami S. K., Rana A., Majumdar S. S., Kumar V., Sarkar D. P. 2001; Sustained activation of mitogen-activated protein kinases and activator protein 1 by the hepatitis B virus X protein in mouse hepatocytes in vivo. J Virol 75:10348–10358 [CrossRef][PubMed]
    [Google Scholar]
  29. Ogden S. K., Lee K. C., Barton M. C. 2000; Hepatitis B viral transactivator HBx alleviates p53-mediated repression of alpha-fetoprotein gene expression. J Biol Chem 275:27806–27814[PubMed]
    [Google Scholar]
  30. Quétier I., Brezillon N., Duriez M., Massinet H., Giang E., Ahodantin J., Lamant C., Brunelle M. N., Soussan P., Kremsdorf D. 2013; Hepatitis B virus HBx protein impairs liver regeneration through enhanced expression of IL-6 in transgenic mice. J Hepatol 59:285–291 [CrossRef][PubMed]
    [Google Scholar]
  31. Santos N. P., Pereira I. C., Pires M. J., Lopes C., Andrade R., Oliveira M. M., Colaço A., Peixoto F., Oliveira P. A. 2012; Histology, bioenergetics and oxidative stress in mouse liver exposed to N-diethylnitrosamine. In Vivo 26:921–929[PubMed]
    [Google Scholar]
  32. Schmittgen T. D., Livak K. J. 2008; Analyzing real-time PCR data by the comparative C T method. Nat Protoc 3:1101–1108 [CrossRef][PubMed]
    [Google Scholar]
  33. Schneider C., Teufel A., Yevsa T., Staib F., Hohmeyer A., Walenda G., Zimmermann H. W., Vucur M., Huss S. other authors 2012; Adaptive immunity suppresses formation and progression of diethylnitrosamine-induced liver cancer. Gut 61:1733–1743 [CrossRef][PubMed]
    [Google Scholar]
  34. Seki E., Brenner D. A., Karin M. 2012; A liver full of JNK: signaling in regulation of cell function and disease pathogenesis, and clinical approaches. Gastroenterology 143:307–320 [CrossRef][PubMed]
    [Google Scholar]
  35. Shirakata Y., Koike K. 2003; Hepatitis B virus X protein induces cell death by causing loss of mitochondrial membrane potential. J Biol Chem 278:22071–22078 [CrossRef][PubMed]
    [Google Scholar]
  36. Sirma H., Giannini C., Poussin K., Paterlini P., Kremsdorf D., Bréchot C. 1999; Hepatitis B virus X mutants, present in hepatocellular carcinoma tissue abrogate both the antiproliferative and transactivation effects of HBx. Oncogene 18:4848–4859 [CrossRef][PubMed]
    [Google Scholar]
  37. Sung W. K., Zheng H., Li S., Chen R., Liu X., Li Y., Lee N. P., Lee W. H., Ariyaratne P. N. other authors 2012; Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat Genet 44:765–769 [CrossRef][PubMed]
    [Google Scholar]
  38. Sze K. M., Chu G. K., Lee J. M., Ng I. O. 2013; C-terminal truncated hepatitis B virus x protein is associated with metastasis and enhances invasiveness by C-Jun/matrix metalloproteinase protein 10 activation in hepatocellular carcinoma. Hepatology 57:131–139 [CrossRef][PubMed]
    [Google Scholar]
  39. Tanaka Y., Kanai F., Ichimura T., Tateishi K., Asaoka Y., Guleng B., Jazag A., Ohta M., Imamura J. other authors 2006; The hepatitis B virus X protein enhances AP-1 activation through interaction with Jab1. Oncogene 25:633–642[PubMed] [CrossRef]
    [Google Scholar]
  40. Toh S. T., Jin Y., Liu L., Wang J., Babrzadeh F., Gharizadeh B., Ronaghi M., Toh H. C., Chow P. K. other authors 2013; Deep sequencing of the hepatitis B virus in hepatocellular carcinoma patients reveals enriched integration events, structural alterations and sequence variations. Carcinogenesis 34:787–798 [CrossRef][PubMed]
    [Google Scholar]
  41. Tu H., Bonura C., Giannini C., Mouly H., Soussan P., Kew M., Paterlini-Bréchot P., Bréchot C., Kremsdorf D. 2001; Biological impact of natural COOH-terminal deletions of hepatitis B virus X protein in hepatocellular carcinoma tissues. Cancer Res 61:7803–7810[PubMed]
    [Google Scholar]
  42. Verna L., Whysner J., Williams G. M. 1996; N-nitrosodiethylamine mechanistic data and risk assessment: bioactivation, DNA-adduct formation, mutagenicity, and tumor initiation. Pharmacol Ther 71:57–81 [CrossRef][PubMed]
    [Google Scholar]
  43. Waris G., Huh K. W., Siddiqui A. 2001; Mitochondrially associated hepatitis B virus X protein constitutively activates transcription factors STAT-3 and NF-κB via oxidative stress. Mol Cell Biol 21:7721–7730 [CrossRef][PubMed]
    [Google Scholar]
  44. Wei C., Ni C., Song T., Liu Y., Yang X., Zheng Z., Jia Y., Yuan Y., Guan K. other authors 2010; The hepatitis B virus X protein disrupts innate immunity by downregulating mitochondrial antiviral signaling protein. J Immunol 185:1158–1168 [CrossRef][PubMed]
    [Google Scholar]
  45. Xia L., Huang W., Tian D., Zhu H., Zhang Y., Hu H., Fan D., Nie Y., Wu K. 2012; Upregulated FoxM1 expression induced by hepatitis B virus X protein promotes tumor metastasis and indicates poor prognosis in hepatitis B virus-related hepatocellular carcinoma. J Hepatol 57:600–612 [CrossRef][PubMed]
    [Google Scholar]
  46. Xiang W. Q., Feng W. F., Ke W., Sun Z., Chen Z., Liu W. 2011; Hepatitis B virus X protein stimulates IL-6 expression in hepatocytes via a MyD88-dependent pathway. J Hepatol 54:26–33 [CrossRef][PubMed]
    [Google Scholar]
  47. Yip W. K., Cheng A. S., Zhu R., Lung R. W., Tsang D. P., Lau S. S., Chen Y., Sung J. G., Lai P. B. other authors 2011; Carboxyl-terminal truncated HBx regulates a distinct microRNA transcription program in hepatocellular carcinoma development. PLoS ONE 6:e22888 [CrossRef][PubMed]
    [Google Scholar]
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