Objective To explore effect of siRNA silencing B-cell specific Moloney murine leukemiavirus integration site (Bmi-1) gene expression on cell proliferation of cervical cancer cells. Methods Hela cells were transferred with control and siRNA Bmi-1. The expression of Bmi-1 in Hela cell was examed by Realtime PCR and western blot. The cell viability was detected by MTT. The cell apoptosis was examed by Annexin V-PI flow cytometry and Hoechst staining. Cell cycle was detected by flow cytometry. The expression of p16, p53, CyclinD1 and Rb was detected by western blot. Results Compared with control group, the expression of Bmi-1 protein and mRNA was down-regulated, the cell viability was decreased, cell apoptotic rate was increased, Cell cycle was arrested in the G1 phase, the expression of p16 was up-regulated, the expresson of p53 and CyclinD1 down-regulated, the phosphorylation of Rb was inhibited in siRNA Bmi-1 group. The difference was statistically significant (P<0.01). Conclusion Bmi-1 inhibited Hela cell proliferation via regulation of expression of cell cycle related protein.