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Objective:To construct a HepG2 cell line which stably expressing Hepatitis B e antigen (HBeAg) and investigate mthe effects of HBeAg on the proliferation,migration and invasion of HepG2 cells.Methods:The lentivirus carrying HBeAg gene was constructed and packaged.HepG2 cells were infected with the lentivirus and screened with puromycin to obtain HepG2 cells which stably expressing HBeAg (HepG2-HBeAg cells).The expression levels of HBeAg mRNA and protein were detected by RT-qPCR and Western blot,respectively.The content of HBeAg secretion in cell supernatant in both HepG2-HBeAg cells and control cells (HepG2-NC cells and HepG2 cells) were detected by IFMA assay.Furthermore,CCK-8 proliferation assay,colony formation assay and transwell migration and invasion assays were conducted to compare the abilities of cell proliferation,migration and invasion,respectively.Results:The expression of HBeAg in the HepG2-HBeAg cell was significantly higher than those in HepG2 cells and HepG2-NC cells.Secreted HBeAg in the supernatant of HepG2-HBeAg cells was 26.33 ± 2.13 PEIU/mL but was undetectable in supernatant of control cells.The proliferation,migration and invasion were all significantly lower in HepG2-HBeAg cells compared to control cells (P＜0.01).Conclusion:A HepG2 cell line which stably expressing HBeAg was constructed successfully,and the over-expression of HBeAg could attenuate the proliferation,migration and invasion of hepatoeellular carcinoma cells.