Experimental study on the effects of miR?671?3p on proliferation and apoptosis of glioma cells

LEI Yi; DENG Teng; CHEN Hainan; HUANG Qianrong; MO Ligen

doi:10.3969/j.issn.1009-0460.2019.09.005

Abstract：Objective To investigate the effect of the expression of miR?671?3p on the proliferation and apoptosis of glioma cells and its possible mechanism. Methods Glioma cell lines U87, U251, U118, A172 and normal human astrocyte HA were cul?tured in vitro. Real?time fluorescence quantitative PCR (QPCR) was used to detect the expression of miR?671?3p in cells. MiR?671?3p siRNA was transfected into U87 cells. CCK?8 assay and cell cloning assay were used to detect the proliferation activity of U87 cells. Flow cytometry was used to detect the changes of cell apoptosis.The relationship between RHOU and miR?671?3p was validated by dual luciferase reporter assay. Results The expression levels miR?671?3p in glioma cells U87, U251, U118 and A172 were 4. 21±0. 18, 3. 78±0. 16, 2. 31±0. 16 and 2. 46±0. 17, respectively, which were significantly higher than those in HA cells (1. 00 ±0. 14), with statistical significance (P＜0. 05). The relative expression of miR?671?3p in miR?671?3p siRNA group was 0. 43± 0. 11, which was significantly lower than that in the control group and the negative control group (P＜0. 05). After 24, 48, 72 and 96 hours of culture, the A values of miR?671?3p siRNA group were 0. 461±0. 085, 0. 524±0. 098, 0. 658±0. 109 and 0. 825±0. 117, which were signifi?cantly lower than those of control group and negative control group (P＜0. 05).The number of colony formation of U87 cells in the miR?671?3p siRNA group was 378±98, which was less than that in the control group (792±92) and the negative control group (750±93), and the difference was statistically significant (P＜0. 05).The apoptotic rate of U87 cells in the miR?671?3p siRNA group was (27. 13± 5. 78)%, which was higher than that in the control group and the negative control group (P＜0. 05). MiR?671?3p mimics inhibited the luciferase activity of wild?type RHOU 3’?UTR, but did not affect the mut?type ( P＞0. 05). Conclusion MiR?671?3p is highly ex?pressed in glioma cells. Downregulated its expression can inhibit the proliferation of glioma cells, increase cell cycle arrest and induce apoptosis, which may be related to RHOU.