Effect of microRNA-96 on proliferation, apoptosis and expression of spindlin1 in cervical cancer cell line HeLa

ZHAO Liang; XIA Chunjun; WU Tangbing; GU Wenlong; XIAO Yanhua

doi:10.3969/j.issn.1009-0460.2017.10.002

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.