miR-509-3p增强卵巢癌细胞对顺铂的敏感性的作用及其机制  

Effect of miR-509-3p on cisplatin sensitivity of ovarian cancer cells and its mechanism

作　　者：武红[1] 张曦辉 王军 张占薪[1] 雷娜[1] WU Hong;ZHANG Xihui;WANG Jun;ZHANG Zhanxin;LEI Na(Department of Gynecology, Zhengzhou People′s Hospital, Zhengzhou 450003, China)

机构地区：[1]郑州人民医院妇科,河南郑州450003

出　　处：《广东药科大学学报》2020年第6期858-863,共6页Journal of Guangdong Pharmaceutical University

基　　金：2018年度河南省医学科技攻关计划项目(2018020731)。

摘　　要：目的探究miR-509-3p对人卵巢癌顺铂耐药SKOV3/DDP细胞的影响。方法RT-PCR检测miR-509-3p、FOXM1在卵巢癌细胞SKOV3和顺铂耐药细胞株SKOV3/DDP中的表达;将SKOV3/DDP细胞转染miR-509-3p mimic,SKOV3细胞转染miR-509-3p inhibitor,RT-PCR检测miR-509-3p表达,蛋白免疫印迹检测FOXM1蛋白表达水平,CCK-8检测细胞活力,流式细胞仪检测细胞凋亡;荧光素酶报告实验验证FOXM1与miR-509-3p靶向关系,构建FOXM1 pcDNA载体过表达FOXM1转染至SKOV3/DDP细胞将细胞分4组:Control组、miR-509-3p组、mimic+pcDNA组和mimic+FOXM1组,蛋白免疫印迹检测FOXM1蛋白表达水平,CCK-8检测细胞活力,流式细胞仪检测细胞凋亡。结果与SKOV3细胞比较,SKOV3/DDP细胞miR-509-3p水平降低,FOXM1水平升高,差异具有统计学意义(P<0.05);在SKOV3/DDP细胞中,与Control组比较,miR-509-3p mimic组miR-509-3p水平升高,FOXM1水平降低,细胞存活率降低,细胞凋亡率升高,差异具有统计学意义(P<0.05);在SKOV3细胞中,与Control组比较,miR-509-3p inhibitor组miR-509-3p水平降低,FOXM1水平升高,细胞存活率升高,细胞凋亡率降低,差异具有统计学意义(P<0.05)。FOXM1与miR-509-3p存在直接靶向作用关系;与Control组比较,miR-509-3p mimic组FOXM1蛋白水平降低,细胞存活率降低,细胞凋亡率降低。与mimic+pcDNA组比较,mimic+FOXM1组FOXM1蛋白水平升高,细胞存活率升高,细胞凋亡率升高,差异具有统计学意义(P<0.05)。结论miR-509-3p通过靶向FOXM1增强卵巢癌细胞对顺铂的敏感性。Objective To investigate the effect of miR-509-3p on cisplatin resistant SKOV3/DDP ovarian cancer cells.Methods RT-PCR was used to detect the expression of miR-509-3p and FOXM1 in SKOV3 cells and cisplatin resistant SKOV3/DDP cells.SKOV3/DDP cells were transfected with miR-509-3p mimic,and SKOV3 cells were transfected with miR-509-3p inhibitor.The expression of miR-509-3p was detected by RT-PCR and the expression of FOXM1 protein was detected by western blot.Cell viability was detected by CCK-8 and cell apoptosis was detected by flow cytometry.Luciferase reporting experiment was used to verify the targeting relationship between FOXM1 and miR-509-3p.The FOXM1 pcDNA vector was transfected into SKOV3/DDP cells,and the cells were divided into four groups including the control group,miR-509-3p group,mimic+pcDNA group and mimic+FOXM1 group.The expression of FOXM1 protein was detected by western blot.Cell viability was detected by CCK-8 and cell apoptosis was detected by flow cytometry.Results Compared with SKOV3 cells,miR-509-3p level was decreased and FOXM1 level was increased in SKOV3/DDP cells(P<0.05).In SKOV3/DDP cells,mir-509-3P mimic group displayed increased miR-509-3p level and cell apoptosis rate,but decreased FOXM1 level and cell survival rate when compared with the control group(P<0.05).In SKOV3 cells,miR-509-3p inhibitor group displayed decreased miR-509-3p level and cell apoptosis rate,but increased FOXM1 level and cell survival rate when compared with the control group(P<0.05).FOXM1 had a direct targeting relationship with miR-509-3p.Compared with the control group,the levels of FOXM1 protein,cell survival rate and apoptosis rate were reduced in the miR-509-3p mimic group.Compared with the mimic+pcDNA group,the levels of FOXM1 protein,cell survival rate and apoptosis rate were increased in the mimic+FOXM1 group(P<0.05).Conclusion miR-509-3p enhances the sensitivity of ovarian cancer cells to cisplatin by targeting FOXM1.

关 键 词：卵巢癌 miR-509-3p 叉头蛋白转录因子M1 多药耐药 

分 类 号：R737.31[医药卫生—肿瘤]