Effects of microRNA-20a on the proliferation and apoptosis of bladder cancer T24 cell

GUO Junsheng; XIA Haibo; MEN Chao; WANG Chunhui; ZHANG Wei

doi:10.3969/j.issn.1009-0460.2018.02.005

Abstract：Objective To investigate the expression of microRNA-20a (miR-20a) in bladder cancer tissues and its effect on the proliferation and apoptosis of human bladder cancer cells. Methods From November 2015 to October 2016, 65 pairs of bladder cancer tissues and para-cancerous tissues were collected in our hospital. Real-time fluorescence quantitative PCR (QPCR) was used to detect the level of miR-20a in cancer tissues and para-cancerous tissues. Lipofectamine(TM) 2000 liposome was used to transfect miR-20a mimics (mimics group) and inhibitor (inhibitor group) into human bladder cancer T24 cells, and T24 cells without transfection were used as control group. The level of miR-20a at 48 h after transfection was detected by QPCR. The proliferation and apoptosis of each group were detected by CCK-8 and Annexin V-FITC/PI double staining via flow cytometry. The target genes of miR-20a were predicted by bioinformatics software Targetscan,and function annotation of the target genes was carried out for Gene Oncology (GO) by cyto-scape3.5.1 and its plug-in CluGO. Results The expression of miR-20a in bladder cancer tissues was 3.181±1.669, which was higher than 1.198±0.756 of the corresponding para-cancerous tissues (P<0.05). The miR-20a levels were 1.024±0.468,2.756±0.941 and 0.412±0.075 at 48 h after transfection in the control group, mimics group and inhibitor group. Compared with the control group, the level of miR-20a increased in the mimics group and decreased in inhibitor group, and the difference was statistically significant (P<0.05). Compared with the control group,the absorbance and proliferative ratio of mimics group increased at 24, 48 and 72 h after transfection, while the absorbance and proliferative ratio of inhibitor group decreased (P<0.05). The apoptotic rates were (12.16±1.67)％,(7.22±0.97)％ and (36.13±4.15)％ at 48 h after transfection in control group, mimics group and inhibitor group, respectively. Compared with the control group, the apoptotic rates decreased in mimics group but increased in the inhibitor group, and the difference was statistically significant (P<0.05). One hundred and forty-seven target genes were screened for miR-20a. The function of GO was mainly enriched in cell cycle, phosphatidyl inositol Akt signaling pathway and cell metabolic process. Conclusion MiR-20a is upregulated in bladder cancer tissue, and it can promote the proliferation of cancer cells and plays a similar role as oncogene. It is expected to become a new target for biological treatment of bladder cancer.