Objective To explore the effect of siRNA targeting the CXCR4 gene on cell proliferation, apoptosis and migration of T24 cells. Methods T24 cells were divided into three groups: interference group, negative control group, and blank control group. Small interfering RNA(siRNA)was transfected together with Lipofectamine 2000 in T24 human bladder cancer cells to block the CXCR4 signal pathway. Western-blot analysis was used to confirm the successful suppression of CXCR4 protein by siRNA. The effect of siRNA on cell death, proliferation and invasiveness was assessed by MTT colorimetric method, cell counting kit-8 assay; the effects of apoptosis was detected by FCM; and Transwell migration assay was performed. Results Expression of CXCR4 protein was successfully suppressed after transfected with siRNA which was verified by Western-blot(P<0.05). Compared with blank control and negative control, the proliferation of T24 bladder cancer cells was successfully restrained and the cell death of T24 bladder cancer cells was significantly promoted by inhibition of CXCR4 gene(all P value<0.05). Transwell test result demonstrated that migration of T24 bladder cancer cells was significantly inhibited after CXCR4 gene was interfered by siRNA(P<0.05). Conclusion This study showed interfering CXCR4 expression level can inhibit cell proliferation, promote cell apoptosis and decreased cell migration ability, which suggested that CXCR4 gene was an important enhancer for the proliferation and migration of bladder cancer cells.