Wnt/β-catenin signaling contributes to diverse cellular functions, such as Drosophila wing development and colon carcinogenesis. Recently, stabilizing mutations of β-catenin, a hallmark of Wnt signaling, were documented in significant numbers of prim ...
Wnt/β-catenin signaling contributes to diverse cellular functions, such as Drosophila wing development and colon carcinogenesis. Recently, stabilizing mutations of β-catenin, a hallmark of Wnt signaling, were documented in significant numbers of primary hepatocellular carcinomas (HCC). However, it has not been established whether the β-catenin mutation leads to the activation of Wnt/β-catenin signaling in hepatoma cells. I found that Wnt/β-catenin signaling could be activated by ectopic expression of Wnt-1 in some hepatoma cells, such as Hep3B and PLC/PRF/5 cells, but not in others, such as Huh7 and Chang cells. Importantly, I noted that the former were derived from hepatitis B virus (HBV)-infected livers, whereas the latter were derived from HBV-negative livers. It was then speculated that HBx, a viral regulatory protein of HBV, is involved in activating Wnt/β-catenin signaling in hepatoma cells. In agreement with this notion, ectopic expression of HBx along with Wnt-1 activated Wnt/β-catenin signaling in Huh7 cells by stabilizing cytoplasmic β-catenin. Further, I showed that such stabilization of β-catenin by HBx was achieved by suppressing glycogen synthase kinase 3 activity via the activation of Src kinase. In conclusion, the data suggested that Wnt-1 is necessary but insufficient to activate Wnt/β-catenin signaling in hepatoma cells and the enhanced stabilization of β-catenin by HBx, in addition to Wnt-1, is essential for the activation of Wnt/β-catenin signaling in hepatoma cells.
HBx, a small regulatory protein of hepatitis B virus, is essential for viral replication. HBx acts as a general transcriptional transactivator in nucleus, whereas it induces multiple signaling pathways in cytoplasm. Stimulation of the viral genome replication by HBx represents one of the most important functions of HBx during infection cycle. A recent study linked the stimulation by HBx to hyperphosphorylation of core protein that leads to the enhanced encapsidation and genome synthesis. In contrast, others attributed the stimulation of the viral genome replication by HBx to its nuclear function largely acting at transcription level. To clarify this discrepancy, I asked what extent nuclear or cytoplasmic HBx contributes to the genome replication. I first attempted to relate its expression level to its subcellular localization. Intriguingly, I found that its abundance primarily affects its subcellular localization in that it is confined to the nucleus at lower abundance, whereas it is predominantly detected in the cytoplasm when overexpressed. Further, I found that HBx, which expressed from endogenous viral promoter in a replicon context, was predominantly localized to nucleus. This finding is reinforced by the analysis of retargeting variants of HBx that harbor either nuclear localization signal (NLS) or nuclear export signal (NES) at its N-terminus. Importantly, NLS-HBx supported the viral genome replication, whereas cytoplasmically localized NES-HBx has impaired ability to support the viral genome replication. The data presented here led me to conclude that nuclear HBx, rather than cytoplasmic HBx, is linked to the stimulation of the viral genome replication.