Abstract: Objective To investigate the effects of toll-like receptor 4 expression on gene expression and cell growth of human HBV-related hepatocellular carcinoma(HCC) cells. Methods Toll-like receptor 4 protein expression were analyzed in five HCC cell lines, Hep3B, HepG2.2.15, HepG2, SMMC7721 and Huh7, by Western blot. The cells with TLR4 overexpression were selected to be further researched. Four groups, normal group, negative control group, plasmid No.3 group and plasmid No.4 group, were divided. After transfecting the HCC cells with TLR4 overexpression via miR-TLR4 plasimids, cell proliferation was measured by MTT assay; cell clonogenicity was detected by clone formation assay; and cell apoptosis and cycle distribution were examined by flow cytometry. Results TLR4 was experssed in all cell lines, and it was the highest in Hep3B cells. Compared with the normal and negative control groups, interfering TLR4 expression could cause cell cycle block at G2/M phase, inhibit the proliferation and cloning efficiency of Hep3B, and promote apoptosis(P＜0.05). Conclusion Up-regulation of TLR4 expression promotes the growth of HBV-related HCC cell Hep3B and cell cycle redistribution, and inhibits cell apotosis, which plays a key role in the carcinogenesis of HBV-related HCC.