Abstract: Objective To investigate the effect of microRNA-96 （miR-96） on proliferation， apoptosis and expression of spindlin1 （SPIN1） in cervical cancer cell line HeLa. Methods The miR-96 analog（mimics） and negative control were transfected into cervical cancer HeLa cells （miR-96 transfection group and miR-96 control group） by liposome， and the HeLa cells without transfection were chosen as non-transfection group. Real time quantitative PCR （QPCR） was used to detect the expression of miR-96 in each group at 48 h after transfection. MTT and flow cytometry were used to detect the proliferative and apoptotic rates at 48 h after transfection in each group. The mRNA and protein levels of SPIN1 in each group were detected by QPCR and Western blotting at 48 h after transfection，respectively. The relationship between miR-96 and SPIN1 was verified by double luciferase target test. Results At 48 h after transfection， the miR-96 levels were 1.068±0.053，1.175±0.084 and 2.434±0.088 in non-transfection group， miR-96 control group and miR-96 transfection group. The miR-96 level in miR-96 transfection group was higher than those of other two groups （P<0.05）. The proliferative rates were （99.633±5.059）%， （97.727±3.079）% and （62.023±5.425）%， and the apoptotic rates were （7.515±0.924）%，（8.123±1.247）% and （26.845±4.126）% in non-transfection group， miR-96 control group and miR-96 transfection group， respectively. Compared with other two groups，the proliferative rate of miR-96 transfection group decreased and the apoptotic rate increased （P<0.05）. The mRNA levels of SPIN1 were 0.965±0.046， 0.917±0.044 and 0.549±0.039，and the protein levels of SPIN1 were 0.667±0.042， 0.715±0.045 and 0.384±0.038 in non-transfection group，miR-96 control group and miR-96 transfection group， respectively. The mRNA and protein levels of SPIN1 in miR-96 transfection group were lower than those of other two groups，and the difference was statistically significant （P<0.05）. MiR-96 inhibited luciferase activity of cells with wild-type SPIN1-3’UTR plasmid， but had no effect on luciferase activity in cells with mutant plasmid. Conclusion Overexpression of miR-96 can inhibit the proliferation and apoptosis of cervical cancer cells and reduce the expression level of SPIN1. MiR-96 can target SPIN1 effectively and can be used as an effective target for the treatment of cervical cancer.