miR-21 influences growth of glioma cells by targeting FasL gene / 中国病理生理杂志

ABSTRACT

AIM:ToinvestigatetheregulationofmiR-21onFasLexpressionanditseffectonthegrowthand apoptosis in glioma cells , and to evaluate the molecular mechanism .METHODS:Differential expression levels of miR-21 in human glioma U251 cells were achieved by transfecting with miR-21 mimics, miR-21 inhibitor or scramble .The viability and apoptosis of U251 cells were detected by CCK-8 assay and flow cytometry with Annexin V-FITC/PI double staining. The recombination vector pmirGLO-FasL was constructed .Dual-luciferase reporter experiment was performed to validate the target genes of miR-21.The expression vector pcDNA3.1-FasL was also constructed , and the biological activity and regula-tory role of miR-21 in U251 cell apoptosis were analyzed by a restore experiment .RESULTS:Exogenous overexpression of miR-21 increased the viability and decreased the apoptosis of U 251 cells ( P<0.05 ) , while miR-21 inhibitors generated the opposite results (P<0.05).Dual-luciferase reporter assay and restore experiment revealed that miR-21 negatively reg-ulated the expression of FasL gene which was regarded as the target gene , thus decreasing the apoptosis of U 251 cells. CONCLUSION:miR-21 increases the viability of glioma U251 cells, in which FasL may be one of the target genes .