Hepatitis B Virus X Protein Inhibits Transforming Growth Factor-β-induced Apoptosis through the Activation of Phosphatidylinositol 3-Kinase Pathway

Transforming growth factor-β (TGF-β) is a potent inducer of apoptosis in Hep 3B cells. This work investigated how hepatitis B virus X protein (HBx) affects TGF-β-induced apoptosis. Trypan blue exclusion and colony formation assays revealed that HBx increased the ID50 toward TGF-β. In the presence of HBx, TGF-β-induced DNA laddering was decreased, indicating that HBx had the ability to block TGF-β-induced apoptosis. Furthermore, HBx did not alter the expression levels of type I and type II TGF-β receptors. HBx did not affect TGF-β-induced activation of promoter activities of the plasminogen activator inhibitor-1 (PAI-1) gene. These results indicate that HBx interferes with only a subset of TGF-β activity. In the presence of phosphatidylinositol (PI) 3-kinase inhibitors, wortmannin or LY294002, the HBx-mediated inhibitory effect on TGF-β-induced apoptosis was alleviated. In addition, the tyrosine phosphorylation levels of the regulatory subunit p85 of phosphatidylinositol 3-kinase (PI 3-kinase) and PI 3-kinase activity were elevated in stable clones with HBx expression. Transactivation-deficient mutants of HBx lost their ability to inhibit TGF-β-induced apoptosis. Phosphorylation of the p85 subunit of PI 3-kinase and Akt, a downstream target of PI 3-kinase, was not observed in stable clones with transactivation-deficient HBx mutant's expression. Thus, the anti-apoptotic effect of HBx against TGF-β can be mediated through the activation of the PI 3-kinase signaling pathway, and the transactivation function of HBx is required for its anti-apoptosis activity.