Hepatitis C virus core promotes hepatic cancer stem cell formation via β-catenin-mediated EpCAM upregulation

Hepatitis C virus (HCV) infection is a major cause of hepatocellular carcinoma (HCC). However, how HCV promotes hepatic progenitor cell (HPC) differentiation into hepatic cancer stem cells (HCSCs) remains unclear, particularly the role of β-catenin signaling and epithelial cell adhesion molecule (EpCAM) regulation in this process. We used mouse HPCs (HP14.5 and HP14.5-Core) and human HCC cell lines (Huh7, HepG2) to investigate HCV core function. Cells were transduced with AdCore, AdGFP, or shRNAs targeting EpCAM or β-catenin. Functional properties were evaluated using spheroid formation, ICG uptake, colony formation, migration, invasion, and in vivo tumorigenesis assays. EpCAM promoter activity and Wnt/β-catenin signaling were examined by luciferase reporter assays. Protein expression was measured by RT-PCR and Western blotting, while immunofluorescence and co-immunoprecipitation were used to analyze HCV core-β-catenin interactions. HCV core expression increased spheroid number, reduced ICG uptake, and accelerated tumor growth in xenograft and orthotopic mouse models, consistent with HPC differentiation toward HCSCs. EpCAM, CD133, CD44, and CD90 were strongly upregulated, whereas EpCAM silencing suppressed proliferation, migration, and invasion. Reporter assays revealed that HCV core enhanced EpCAM promoter activity and activated Wnt/β-catenin signaling. Immunofluorescence demonstrated β-catenin nuclear translocation, and co-immunoprecipitation confirmed direct interaction between HCV core and β-catenin. Knockdown of β-catenin reduced EpCAM, c-Myc, and cyclin D1 expression, diminished spheroid formation, restored ICG uptake, and significantly impaired tumorigenesis in vivo. The HCV core-β-catenin-EpCAM axis drives the conversion of HPCs into HCSCs. This pathway is a central mechanism in HCV-related HCC.IMPORTANCEHCV core protein directly interacts with β-catenin, driving its nuclear translocation and activating EpCAM expression. This promotes HPC differentiation into HCSCs, marked by increased spheroid formation, migration, invasion, and tumorigenesis. Silencing EpCAM or β-catenin reverses these effects, confirming that the HCV core-β-catenin-EpCAM axis is a key pathway in HCV-related HCC.