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

李坤; 张晶

doi:10.13898/j.cnki.issn.1000-2200.2022.05.005

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

李坤,
张晶

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

LI Kun,
ZHANG Jing

摘要

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。

Abstract:
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).
MethodsHuman 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.
ResultsCompared 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), and there was no significant difference in luciferase activity between the miR-367 NC+MUT group and miR-367 mimics+MUT group(P>0.05).
ConclusionsOverexpression of miR-367 may promote the proliferation, migration and invasion of endometrial cancer HEC-1A cells by targeted inhibiting TET2.

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