Role Of Wnt/β-catenin Signaling Transduction Pathway In The Study Of Hepatitis B Virus X Gene Enhances Cell Survival Of Oval Cells In Vitro

Objectives To investigate the role of Wnt/β-catenin signaling transduction pathway in the study of Hepatitis B virus X gene enhances cell survival of oval cells in vitro.Methods On the basis of the establishment of pEGFP-HBx-LE/6 cell line with stable transfected HBx gene, pEGFP-LE/6 oval cells and LE/6 oval cells were used as control cells , Real-time PCR, Immunocytochemistry, Immunofluorescence and Western Blot were facilitated to observe the expression ofβ-catenin gene, cyclin D1 gene and c-myc gene before and after the HBx gene transfection in LE/6 oval cells. The changes of morphology, proliferation and cell cycle were studied by Transmission Electron Microscopy and Inversion phase contrast microscope, MTT assay and Flow cytometry, respectively.β-catenin specific siRNA was used to testify the function ofβ-catenin in proliferation of pEGFP-HBx-LE/6 oval cells.Results Compared with control LE/6 oval cells without transfection, the Wnt/β-catenin signaling transduction pathway of pEGFP-HBx-LE/6 oval cells was activated after transfection with HBx gene. The mRNA and protein levels ofβ-catenin gene, cyclin D1 gene and c-myc gene in pEGFP-HBx-LE/6 oval cells rose obviously. The difference was statistical significance(P<0.05).The pEGFP-HBx-LE/6 oval cells showed bigger size with malformed nucleus by Inversion phase contrast microscope and Transmission Electron Microscopy. MTT assay demonstrated that cell growth in pEGFP-HBx-LE/6 oval cells was faster than control groups. Flow cytometry suggested that, compared with the control cells, the pEGFP-HBx-LE/6 oval cells changed cell cycle from G0/G1 phase to S or G2/M phase, especially, the proportion of aneuploid cells increased. After transfection withβ-catenin siRNA, the proliferation of pEGFP-HBx-LE/6 oval cells was significantly decreased(P<0.05).Conclusions Wnt/β-catenin signaling transduction pathway is one of the routes for HBx to enhance cell survival of oval cells in vitro.