摘要
Abstract
Objective:To investigate the impacts of long non-coding RNA KCNQ1 overlapping transcript 1(LncRNA KCNQ1OT1)on proliferation and radiation sensitivity of glioma cells,and its regulatory mechanism on the miR-181a-5p/TRIM71 pathway.Methods:Human glioma cells U251 were divided into NC group(untransfected plasmid),si-NC group(transfected with KCNQ1OT1 negative control),si-KCNQ1OT1 group(transfected with si-KCNQ1OT1),si-KCNQ1OT1+miR-NC group(co-transfected with si-KCNQ1OT1 and miR-181a-5p negative control),si-KCNQ1OT1+miR-181a-5p inhibitor group(co-transfected with si-KCNQ1OT1 and miR-181a-5p inhibitor).After 48 hours of transfection,cells selected from each group were irradiated with 6 Gy radiation.RT-qPCR method was applied to detect the levels of KCNQ1OT1,miR-181a-5p,and TRIM71 mRNA of cells in each group.CCK-8 method and colony formation assay were applied to assess the proliferation activity of transfected cells in each group.Flow cytometry was adopted to detect the apoptosis of transfected cells in each group.Western blotting was performed to measure the relative expression levels of TRIM71 protein in transfected cells of each group.Dual luciferase reporter gene assay was applied to verify the targeting relationship between miR-181a-5p and KCNQ1OT1,TRIM71.Results:Compared with the si-NC group,the expression levels of KCNQ1OT1 and TRIM71 mRNA,cell survival rate,and number of clones formed in the si-KCNQ1OT1 group decreased,while the expression level of mi R-181a-5p increased.After vertical irradiation with 6 Gy of radiation,the survival rate of transfected cells in all groups decreased,while the apoptosis rate increased.Among them,the survival rate of cells in the si-KCNQ1OT1+6Gy group was lower than that in the si-NC+6Gy group,while the apoptosis rate was higher in the si-NC+6Gy group.The rescue experiment demonstrated that miR-181a-5p inhibitor reversed the effects of KCNQ1OT1 knockdown on cell proliferation and radiotherapy sensitivity.Dual luciferase activity confirmed the targeting relationship between miR-181a-5p and KCNQ1OT1,or TRIM71.Conclusion:KCNQ1OT1 knockdown can inhibit the proliferation of glioma cells and improve their radiotherapy sensitivity by up-regulating miR-181a-5p and down-regulating TRIM71 expression.关键词
胶质瘤/长链非编码RNA KCNQ1重叠转录物1/miR-181a-5p/TRIM71Key words
glioma/long non-coding RNA KCNQ1 overlapping transcript 1/miR-181a-5p/TRIM71分类
医药卫生
