ObjectiveTo evaluate effect of hepatitis B virus (HBV) on hepatocyte senescence through HBV X protein (HBx). MethodsHepG2.2.15 cells, HepG2.2.15  cells treated by HBx siRNA (HepG2.2.15HBxsi), and HepG2 cells with overexpressed HBx (HepG2HBx) were performed βgalactosidase staining, hepatocyte senescenceassociated protein was detected by Western Blot; effect of HBx on hepatocyte senescence was compared. ResultsThe proportions of positive cells for βgalactosidase staining in each group were as follows: (0.480±0.096) in HepG2.2.15 group, (0.016±0.005) in HepG2 group, difference between two groups was statistically significant (P<0.001); (0.278±0.065) in HepG2.2.15HBxsi group, (0.329±0.044) in its’ control group（HepG2.2.15HBxnc group)， difference between two groups was not statistically significant(P=0.092）; (0.319±0.033) in HepG2HBx group, (0.064±0.012) in it’s control group(HepG2HBxcon group), difference between two groups was statistically significant(P<0.001). Western Blot showed that the expression of hepatocyte senescenceassociated protein p53 and p21 in HepG2.2.15 group and HepG2HBx group were significantly higher than those of control group, while p53 and p21 in HepG2.2.15HBxsi group were significantly lower than that of HepG2.2.15HBxnc group. ConclusionHBV can promote hepatocellular senescence through HBx protein.