circMAN2B2对宫颈癌细胞顺铂敏感性的影响  

Effects of circMAN2B2 on the sensitivity of cervical cancer cells to cisplatin

作　　者：蒙秋[1] 谢兴润 张静[1] 王海燕[1] 黄守国[1] MENG Qiu;XIE Xingrun;ZHANG Jing(Department of Obstetrics and Gynecology,Haikou Hospital of Xiangya Medical College of Central South University,Hainan,Haikou 570208,China;不详)

机构地区：[1]中南大学湘雅医学院附属海口医院妇产科,海口市570208 [2]中南大学湘雅医学院附属海口医院放射科,海口市570208

出　　处：《河北医药》2021年第22期3376-3379,3384,共5页Hebei Medical Journal

摘　　要：目的探讨circMAN2B2对宫颈癌细胞顺铂敏感性的影响及分子机制。方法采用顺铂梯度暴露法构建顺铂耐药宫颈癌细胞株HeLa/DDP,将其分为DDP+si-NC组、DDP+si-circMAN2B2组、DDP+si-circMAN2B2+anti-miR-NC组、DDP+si-circMAN2B2+anti-miR-216a-3p组。四甲基偶氮唑盐比色法(MTT)检测细胞增殖抑制率及半数抑制浓度(IC50);实时荧光定量PCR(RT-qPCR)检测circMAN2B2和miR-216a-3p的表达水平;双荧光素酶报告实验检测circMAN2B2和miR-216a-3p的靶向调控;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测蛋白表达。结果与HeLa细胞比较,HeLa/DDP细胞增殖抑制率显著降低,IC50值升高,circMAN2B2表达水平升高,miR-216a-3p表达水平降低,差异均有统计学意义(P<0.05)。StarBase预测显示,circMAN2B2的序列中含有与miR-216a-3p互补的核苷酸序列。WT-circMAN2B2与miR-216a-3p共转染的细胞荧光素酶活性低于WT-circMAN2B2与miR-NC组共转染的细胞(P<0.05),而MUT-circMAN2B2与miR-216a-3p或miR-NC共转染的细胞荧光素酶活性比较差异无统计学意义。过表达circMAN2B2,miR-216a-3p表达水平降低(P<0.05),抑制circMAN2B2表达后miR-216a-3p表达水平升高(P<0.05)。与DDP+si-NC组比较,DDP+si-circMAN2B2组circMAN2B2表达水平降低,miR-216a-3p表达水平升高,细胞增殖抑制率升高,细胞凋亡率升高,Ki-67、Bcl-2表达水平降低,Bax表达水平升高,差异均有统计学意义(P<0.05)。与DDP+si-circMAN2B2+anti-miR-NC组比较,DDP+si-circMAN2B2+anti-miR-216a-3p组miR-216a-3p表达水平降低,细胞增殖抑制率降低,细胞凋亡率降低,Ki-67、Bcl-2表达水平升高,Bax表达水平降低,差异均有统计学意义(P<0.05)。结论抑制circMAN2B2表达可能通过上调miR-216a-3p增强宫颈癌细胞对顺铂的敏感性。Objective To investigate the effects of circMAN2B2 on the sensitivity of cervical cancer cells to cisplatin and its molecular mechanism.Methods The cisplatin resistant cervical cancer cell line HeLa/DDP was established by cisplatin gradient exposure method,which were divided into DDP+si-NC group,DDP+si-circMAN2B2 group,DDP+si-circMAN2B2+anti miR-NC group,DDP+si-circMAN2B2+anti-miR-216a-3p group.The cell proliferation inhibition rate and half inhibitory concentration(IC50)were detected by Tetramethylazolium salt colorimetric method(MTT),the expression levels of circMAN2B2 and miR-216a-3p were detected by real time fluorescent quantitative PCR(RT-qPCR),the targeted regulation of circMAN2B2 and miR-216a-3p were detected by dual luciferase reporter experiment,the cell apoptosis was detected by flow cytometry experiment,the protein expressions was detected by Western blot.Results As compared with that of HeLa cells,the proliferation inhibition rate of HeLa/DDP cells was significantly decreased,and IC50 value was significantly increased,and the expression levels of circMAN2B2 were significantly increased,but the expression levels of miR 216a-3p were significantly decreased(P<0.05).The StarBase prediction showed that there was nucleotide sequence which was complementary with miR-216a-3p.Moreover the luciferase activity of the cell which was co-transfected by the sequence of WT circMAN2B2 and miR-216a-3p was significantly lower than that of cells which was co-transfected by WT circMAN2B2 and miR NC(P<0.05),however there was no sinificant differences of luciferase activity of the cells between which were co-transfected by MUT circMAN2B2 and miR-216a-3p and which were co-transfected by MUT circMAN2B2 and miR-NC.After over-expression of circMAN2B2,the expression levels of miR-216a-3p were significantly decreased(P<0.05).After inhibiting the expression of circMAN2B2,the expression levels of miR-216a-3p were significantly increased(P<0.05).As compared with those in DDP+si-NC group,in DDP+si-circMAN2B2 group,the expression lev

关 键 词：circMAN2B2 miR-216a-3p 宫颈癌 顺铂 敏感性 

分 类 号：R737.33[医药卫生—肿瘤]