β-catenin is overexpressed in hepatic fibrosis and blockage of Wnt/β-catenin signaling inhibits hepatic stellate cell activation

beta-catenin, a core component of Wnt/beta-catenin signaling, has been shown to be an important regulator of cellular proliferation and differentiation. Abnormal activation of Wnt/beta-catenin signaling promotes tissue fibrogenesis. In the present study, the role of beta-catenin during liver fibrogenesis was analyzed and the functional effects of beta-catenin gene silencing in hepatic stellate cells (HSCs) using small interfering (si)RNA were investigated. The expression of beta-catenin in human hepatic fibrosis tissues of different grades and normal human hepatic tissues was examined using immunohistochemistry. To inhibit the Wnt/beta-catenin signaling pathway, siRNA for beta-catenin was developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000. beta-catenin expression was evaluated by quantitative polymerase chain reaction (qPCR) and western blot analysis. The expression of collagen types I and III was evaluated by qPCR and immunofluorescent staining. Cellular proliferation and the cell cycle were analyzed using a methyl thiazolyl tetrazolium assay. Apoptosis was assessed by Annexin V staining. A higher expression level of beta-catenin was identified in the patients with high-grade hepatic fibrosis in comparison with that of the normal controls. Additionally, beta-catenin siRNA molecules were successfully transfected into HSCs and induced inhibition of beta-catenin expression in a time-dependent manner. beta-catenin siRNA treatment also inhibited synthesis of collagen types I and III in transfected HSCs. Furthermore, compared with those of the control group, siRNA-mediated knockdown of beta-catenin in HSC-T6 cells inhibited cell proliferation and resulted in cell apoptosis. This study suggests a significant functional role for beta-catenin in the development of liver fibrosis and demonstrates that downregulation of the Wnt/beta-catenin signaling pathway inhibits HSC activation. Thus, this study provides a novel strategy for the treatment of hepatic fibrosis.