| ABSTRACT Objective: This study was to investigate the role of LGALS3BP in the IFN-γ mediated anti-HBV process in human hepatocytes in vitro. Methods: HepG2 and HepG2.215 cell lines were transiently transfected with LGALS3BP overexpression plasmid and its interfering plasmid, and levels of LGALS3BP transcription and translation were detected by means of qRT-PCR and Western Blot. CCK-8 was used to determine the optimal concentration of IFN-γ treatment for HepG2 and HepG2.215 cells, and evaluate the cell proliferation. The effects of IFN-γ treatment on transcription and translation of endogenous LGALS3BP in HepG2 cells were investigated using qRT-PCR and Western Blot. When changing LGALS3BP expression, the effect of IFN-γ stimulation on the exocrine HBsAg and HBeAg were examined using ELISA, and levels of DNA in HBV nucleocapsid and intracellular RNA were evaluated using q-PCR and RT-PCR, respectively. Results: After treated with 40 ng/ml IFN-γ for 48 hours, the level of LGALS3BP mRNA in HepG2 cells was 2.87 folds higher than that in the control group, whereas the change of LGALS3BP expression had no significant effect on the cell proliferation. In case that LGALS3BP was overexpressed in HepG2.215 cells during the IFN-γ mediated anti-HBV process, compared with the control group, it was found the relative decrease rates of extracellular HBsAg, HBeAg as well as of the relative decrease rates of intracellular HBV DNA- and RNA levels were 46%, 67%, 59% and 49%, respectively; For HepG2 cells that were transiently transfected with pCH9-3091, compared with the control group, the relative decrease rates of extracellular HBsAg, HBeAg as well as the intracellular viral DNA and RNA were 67%, 53%, 60% and 29%, respectively. In contrast, in case that LGALS3BP was down-regulated by siRNA interfering-plasmid SHLG, in the process of IFN-γ mediated anti HBV for HepG2.215, compared with the control group, the relative increase rates of secreted HBsAg, HBeAg as well as of intracellular viral DNA- and RNA levels were 46%, 67%, 59% and 49%, respectively; For HepG2 that transiently transfected with pCH9-3091, the relative increase rates of HBsAg, HBeAg as well as of intracellular viral DNA and RNA levels were 67%, 77%, 67% and 45%, respectively. Conclusion: LGALS3BP is a positive regulator and participates in the IFN-γ mediated anti-HBV process. |
