miR-367通过靶向调控TET2对子宫内膜癌细胞增殖、迁移及侵袭的影响  

Effects of miR-367 on proliferation,migration and invasion of endometrial cancer cells by targeted regulating TET2

作　　者：李坤[1] 张晶[2] 

机构地区：[1]河北北方学院附属第一医院妇科门诊,河北张家口075000 [2]河北北方学院附属第一医院妇科,河北张家口075000

出　　处：《蚌埠医学院学报》2022年第5期580-585,共6页Journal of Bengbu Medical College

基　　金：河北省医学科学研究项目(20211818)。

摘　　要：目的:探讨miR-367通过靶向调控10-11异位的甲基胞嘧啶双加氧酶2(TET2)对子宫内膜癌细胞增殖、迁移及侵袭的影响。方法:将人子宫内膜癌HEC-1A细胞分为5组:空白组(NG)、阴性转染组(mimics NC)、miR-367过表达组(miR-367 mimics)、inhibitor NC组、miR-367 inhibitor组。qRT-PCR检测HEC-1A细胞中TET2 mRNA、miR-367表达水平;MTT法检测细胞增殖情况;Transwell法检测细胞侵袭和迁移能力;TargetScan数据库预测miR-367的靶基因并用双荧光素酶报告基因实验加以验证;Western blotting检测TET2、c-myc、cyclin D1、MMP-2、MMP-9蛋白表达水平。结果:与NG、mimics NC组相比,miR-367 mimics组HEC-1A细胞TET2 mRNA表达水平下降(P<0.05),miR-367表达水平升高,24、48 h细胞存活率上升,侵袭、迁移细胞数增加(P<0.05),TET2蛋白表达水平降低(P<0.05),MMP-2、MMP-9、c-myc、cyclin D1蛋白表达水平升高(P<0.05)。与NG、inhibitor NC组相比,miR-367 inhibitor组HEC-1A细胞TET2 mRNA表达水平升高(P<0.05),miR-367表达水平下降,24、48 h细胞存活率降低,侵袭、迁移细胞数减少,TET2蛋白表达水平升高(P<0.05),MMP-2、MMP-9、c-myc、cyclin D1蛋白表达水平降低(P<0.05)。TargetScan数据库预测显示miR-367是TET2潜在靶基因。荧光素酶报告实验结果显示,miR-367 mimics+WT组HEC-1A细胞荧光素酶活性低于miR-367 NC+WT组(P<0.05),miR-367 NC+MUT组与miR-367 mimics+MUT组荧光素酶活性差异无统计学意义(P>0.05)。结论:miR-367过表达可能通过靶向抑制TET2促进子宫内膜癌HEC-1A细胞增殖、迁移和侵袭。Objective:To investigate the effect of miR-367 on the proliferation,migration and invasion of endometrial cancer cells by targeted regulating ten-eleven translocation methylcytosine dioxygenase 2(TET2).Methods:Human endometrial cancer HEC-1A cells were divided into 5 groups:blank group(NG),negative transfection group(mimics NC),miR-367 overexpression group(miR-367 mimics),inhibitor NC group and miR-367 inhibitor group.qRT-PCR was used to determine the expression levels of TET2 mRNA and miR-367 in HEC-1A cells;MTT assay was applied to detect cell proliferation;Transwell assay was carried out to detect cell invasion and migration ability;TargetScan database was employed to predict the target gene of miR-367,and dual luciferase reporter gene assay was used to verify the relationship;Western blotting was performed to analyze the protein expression levels of TET2,c-myc,cyclin D1,MMP-2 and MMP-9.Results:Compared with the NG group and mimics NC group,in the miR-367 mimics group,the expression level of TET2 mRNA in HEC-1A cells decreased(P<0.05),the expression level of miR-367 increased(P<0.05),the cell survival rate aftertreated for 24,48 h and number of invaded and migrated cells increased(P<0.05),the protein expression level of TET2 decreased(P<0.05),the protein expression levels of MMP-2,MMP-9,c-myc and cyclin D1 increased(P<0.05).Compared with NG group and inhibitor NC group,in the miR-367 inhibitor group,the expression level of TET2 mRNA in HEC-1A cells increased(P<0.05),the expression level of miR-367 decreased(P<0.05),the cell survival rate aftertreated for 24,48 h and number of invaded and migrated cells decreased,the protein expression level of TET2 increased(P<0.05),the protein expression level of MMP-2,MMP-9,c-myc and cyclin D1 decreased(P<0.05).TargetScan database prediction showed that miR-367 was a potential target gene of TET2.The results of the luciferase reporter assay showed that the luciferase activity of HEC-1A cells in the miR-367 mimics+WT group was lower than that in the miR-367 NC+WT group(P<0.05

关 键 词：子宫颈肿瘤 miR-367 10-11异位的甲基胞嘧啶双加氧酶2 细胞增殖 迁移 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]