Abstract:To examine the transforming potential of the × gene product of hepatitis B virus (HBV), the X‐gene‐containing region (referred to as the HBx region) was introduced into mouse NIH3T3 cells. Each transformed cell line expressed X‐coding mRNA at a different level. A positive correlation was found between the level of X‐coding mRNA and the saturation density of the cells. The HBx‐transformed cell lines exhibited × protein production and tumor formation in nude mice. The function of HBV in oncogenesis may involve t… Show more
“…However, in a di erent genetic context, HBx transgenic mice show either no obvious pathology (Lee et al, 1990), only increased susceptibility to chemical carcinogens (Slagle et al, 1996), or acceleration of the development of c-mycinduced HCC (Terradillos et al, 1997). Only a few studies have shown transformation of certain cell types by HBx in vitro (Hohne et al, 1990;Shirakata et al, 1989). Thus, the actual impact of HBx in liver carcinogenesis and the mechanisms of its action remain largely obscure.…”
Chronic infection by HBV is the leading cause of hepatocellular carcinoma in man. Several lines of evidence suggest that the viral transactivator HBx plays a critical role in the molecular pathogenesis of HBVrelated HCC. To study the actual impact of HBx and the mechanism of its action, we have recently cloned and characterized a set of X-sequences from HCC in patients with chronic infection by HBV. In the present study, we have compared the e ects of HBx and its naturally arising mutants on cell growth and viability. We report that HBx inhibits clonal outgrowth of cells and induces apoptosis by a p53-independent pathway. Furthermore, HBx expression induced a late G1 cell cycle block prior to their counterselection by apoptosis. Importantly, mutations in the HBx-gene evolving in hepatocellular carcinoma abolished both HBx-induced growth arrest and apoptosis. Using a panel of engineered mutants we have mapped the growth suppressive e ect of HBx to domains shown to be required for its transactivating function. Based on these results, we propose that abrogation of the anti-proliferative and apoptotic e ects of HBx by naturally occurring mutations might render the hepatocytes susceptible to uncontrolled growth and contribute to multistep hepatocarcinogenesis associated with HBV-infection.
“…However, in a di erent genetic context, HBx transgenic mice show either no obvious pathology (Lee et al, 1990), only increased susceptibility to chemical carcinogens (Slagle et al, 1996), or acceleration of the development of c-mycinduced HCC (Terradillos et al, 1997). Only a few studies have shown transformation of certain cell types by HBx in vitro (Hohne et al, 1990;Shirakata et al, 1989). Thus, the actual impact of HBx in liver carcinogenesis and the mechanisms of its action remain largely obscure.…”
Chronic infection by HBV is the leading cause of hepatocellular carcinoma in man. Several lines of evidence suggest that the viral transactivator HBx plays a critical role in the molecular pathogenesis of HBVrelated HCC. To study the actual impact of HBx and the mechanism of its action, we have recently cloned and characterized a set of X-sequences from HCC in patients with chronic infection by HBV. In the present study, we have compared the e ects of HBx and its naturally arising mutants on cell growth and viability. We report that HBx inhibits clonal outgrowth of cells and induces apoptosis by a p53-independent pathway. Furthermore, HBx expression induced a late G1 cell cycle block prior to their counterselection by apoptosis. Importantly, mutations in the HBx-gene evolving in hepatocellular carcinoma abolished both HBx-induced growth arrest and apoptosis. Using a panel of engineered mutants we have mapped the growth suppressive e ect of HBx to domains shown to be required for its transactivating function. Based on these results, we propose that abrogation of the anti-proliferative and apoptotic e ects of HBx by naturally occurring mutations might render the hepatocytes susceptible to uncontrolled growth and contribute to multistep hepatocarcinogenesis associated with HBV-infection.
“…Numerous attempts have been made to examine the oncogenic potential of HBx in cell culture (Hohne et al, 1990;Oguey et al, 1996;Shirakata et al, 1989;Tarn et al, 1999). However, its transforming ability was barely measurable only when cells were immortalized by other oncogenes, such as SV40 T-antigen (Hohne et al, 1990) or TGF-a (Oguey et al, 1996;Tarn et al, 1999).…”
“…The fact that HBxAg and WHxAg are also structurally and functionally similar (Galibert et al, 1982), as are both human and woodchuck p53s, further suggests that they may share common properties, possibly including some of the mechanistic steps whereby these viruses mediate hepatocellular transformation. Given the increasing importance of HBxAg to transformation 1994;Shirakata et al, 1989;Hohne et al, 1990;Koike et al, 1994), combined with its complexing to wild type p53 (Feitelson et al, 1993a;Ueda et al, 1995;Truant et al, 1995), it was of great interest to see whether this also occurred in the WHV system. The results of this study show that WHxAg and woodchuck p53 form complexes both in vitro and in vivo in a manner analogous to that which occurs in human carriers with HCC, providing at least part of a common mechanism whereby these viruses cause hepatocellular carcinoma.…”
Section: Discussionmentioning
confidence: 99%
“…The ®nding that HBxAg is often expressed from viral DNA fragments integrated into human liver or HCC nodules, and that the HBxAg made is still capable of trans-activation (Zahm et al, 1988;Wollersheim et al, 1988), suggest that functional HBxAg is associated with the development of these tumors, although it is not known whether HBxAg associated trans-activation is relevant to hepatocarcinogenesis. In addition, the ®nding that HBxAg transforms NIH3T3 cells (Shirakata et al, 1989) and a nontransformed mouse hepatocyte cell line (Hohne et al, 1990), suggests that under selected conditions, HBxAg can mediate transformation of cultured cells. Further, the ®nding that HBxAg complexes with the tumor suppressor product, p53, in the majority of HBV associated HCC Figure 6 Complex formation between woodchuck p53 and WHxAg in vivo.…”
Section: Discussionmentioning
confidence: 99%
“…For example, HB6Ag expression is sustained at high levels in chronic carriers who develop HCC, and in many cases, it is the only HBV encoded product detected by immunohistochemistry 1991a). Further, HB6Ag causes tumorigenic transformation of NIH3T3 cells (Shirakata et al, 1989), and of an untransformed mouse hepatocyte cell line (Hohne et al, 1990). More recently, HB6Ag has been shown to complex with and inactivate p53 both in vitro and in vivo (Feitelson et al, 1993a;Wang et al, 1994;Truant et al, 1995), suggesting that this relationship may be important for tumor development.…”
Full length cDNAs for p53 were made by reverse transcription-polymerase chain reaction of total RNA from two normal woodchuck livers. Two randomly chosen clones from each liver were sequenced and shown to be identical. This sequence revealed 80% or more identity with p53 sequences from human, monkey, and mouse. The cDNA was translated into a *55 kD protein in vitro that was immunoprecipitated by antibodies to p53. Cotranslation of woodchuck p53 with woodchuck hepatitis virus X antigen, followed by immunoprecipitation suggested X/p53 complex formation. Similar complexes were also immunoprecipitated from extracts of infected liver, but not from uninfected liver. The ®nding of X/p53 complexes in vivo and in vitro in the woodchuck hepadnavirus system, combined with analogous data with hepatitis B, suggests a common mechanism by which these viruses contribute to hepatocellular transformation.
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