Objective To investigate the effects of circular RNA MAPK4(circ-MAPK4)on biological behaviors of glioma cells.Methods The expression level of circ-MAPK4 and miR-125a-3p in human glioma cell lines(U138,U373,U87,A172,U251)and normal human astrocytes(HA1800)was detected by real-time fluorescence quantitative polymerase chain reaction(qPCR).U138 cells were divided into low-expression circ-MAPK4 group(si-circMAPK4 group),negative control group(si-NC group)and non-transfection group(control group).U138 cells in si-circMAPK4 group and si-NC group were transfected with circ-MAPK4 siRNA and its negative control SIRNA-NC,while cells in control group were not transfected.The targeted relationship between circ-MAPK4 and miR-125a-3p was predicted by circNet and StarBase analysis,which was verified by luciferase assay.U138 cells in si-circMAPK4 group were divided into two subgroups(circ-MAPK4+miR-inhibitor group,circ-MAPK4+NC-inhibitor group),and relevant transfection was completed.The activity of U138 cells was detected by CCK-8,formation rate of cell clone was detected by colony-forming assay,number of invasion cells was detected by Transwell assay,apoptosis rate was detected by flow cytometry,and expression levels of cell proliferation proteins(Ki67,cycD),invasion proteins(E-cadherin,N-cadherin,Vimentin)and apoptosis proteins(Ceaved-caspase3/caspase3)were detected by Western blot.Results Compared with HA1800 cells,expression level of circ-MAPK4 increased in U138,U373,U87,and A172 cells(P＜0.05),while expression level of miR-125a-3p decreased(P＜0.05).The results of luciferase assay showed that relative activities of luciferase in circ-MAPK4-mut/miR-125a-3p mimics,circ-MAPK4-mut/NC-mimics and circ-MAPK4-WT/NC-mimics cells were higher than those in circ-MAPK4-WT/miR-125a-3p mimics cells(F=437.659,P＜0.001).Compared with si-NC group,cells activity,clone formation rate,number of invasion cells,expressions of Ki67,cycD,N-cadherin and Vimentin proteins decreased(P＜0.05);while apoptosis rate,expressions of Ceved-caspase3/caspase3 and E-cadherin proteins increased in si-circMAPK4 group(P＜0.05).Compared with circ-MAPK4+NC-inhibitor group,cells activity,clone formation rate,number of invasion cells,expressions of Ki67,cycD,N-cadherin,and Vimentin proteins increased(P＜0.05);while apoptosis rate,expressions of Ceved-caspase3/caspase3 and E-cadherin proteins decreased in circ-MAPK4+miR-inhibitor group(P＜0.05).Conclusion The low-expression circ-MAPK4 could weaken proliferation and invasion of human glioma U138 cells,and promote apoptosis,which might be related to the adsorption of miR-125a-3p.
维普
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