MiR-17-5p在子宫内膜异位症间质细胞中的表达及其对基质金属蛋白酶-2的影响

目的:探讨miR-17-5p在子宫内膜异位症(endometriosis,EMs)间质细胞中的表达及其对基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)的影响。方法:MiRNA芯片筛选EMs异位子宫内膜组织中差异表达的miRNA,qRT-PCR验证EMs异位、在位子宫内膜组织和子宫肌瘤正常子宫内膜组织(对照组)中miR-17-5p的表达;提取以上各组组织中原代间质细胞,免疫荧光鉴定表面标志物;qRT-PCR和蛋白质印迹法检测各组间质细胞中miR-17-5p和MMP-2的表达;双荧光素酶报告基因实验验证miR-17-5p和MMP-2的靶向关系;EMs异位间质细胞中转染miR-17-5p mimic和inhibitor,qRT-PCR、蛋白质印迹法和明胶酶谱法检测MMP-2的表达,克隆形成实验检测细胞增殖能力,Transwel l实验检测细胞侵袭能力,划痕实验检测细胞迁移能力。结果:EMs异位子宫内膜组织中miR-17-5p、miR-200b、miR-106b、miR-15b、miR-141、miR-22显著下降,miR-202、miR-150、miR-365表达显著上升;与对照组相比,miR-17-5p在EMs异位、在位子宫内膜组织和间质细胞中表达均显著下降(P<0.05),而MMP-2在EMs异位、在位子宫内膜间质细胞中表达均显著升高(P<0.05);与阴性对照组(negative control,NC)相比,过表达miR-17-5p抑制MMP-2的表达并显著抑制细胞克隆形成数,细胞侵袭数及细胞迁移率(P<0.05),敲低miR-17-5p,结果相反。结论:MiR-17-5p在EMs间质细胞中表达下降,抑制miR-17-5p的表达能够促进MMP-2表达升高同时增强细胞增殖、侵袭和迁移能力。

Expression of miR-17-5p in endometriosis stromal cells and its effect on matrix metalloproteinase-2

Objective:To investigate the expression of miR-17-5p in endometriosis(EMs)stromal cells and its effect on matrix metalloproteinase-2(MMP-2).Methods:The miRNA chip was used to screen differentially expressed miRNAs in ectopic endometrial tissues of EMs.qRT-PCR was used to verify the expression of miR-17-5p in ectopic,orthotopic endometrial tissue of EMs and normal endometrial tissue of uterine fibroids(control group).Primary mesenchymal cells were extracted from the above groups,and surface markers were identified by immunofluorescence;qRT-PCR and Western blotting were used to detect the expression of miR-17-5p and MMP-2 in mesenchymal cells of each group;dual luciferase reporter assay was used to confirm the targeting relationship between miR-17-5p and MMP-2.EMs were transfected with miR-17-5p mimic and inhibitor,qRT-PCR,Western blot and Zymographic detection were used to detect MMP-2 expression;the colony formation test was used to detect cell proliferation ability,the Transwell assay was used to detect cell invasion ability,and the scratch test was used to detect cell migration ability.Results:The expressions of miR-17-5p,miR-200b,miR-106b,miR-15b,miR-141,and miR-22 in ectopic endometrial tissue decreased significantly,and the expressions of miR-202,miR-150,and miR-365 increased significantly.Compared with the control group,miR-17-5p expression was significantly decreased in EM ectopic,eutopic endometrial tissue and interstitial cells(P<0.05),while the expression of MMP-2 was significantly increased in ectopic and eutopic endometrial stromal cells(P<0.05);compared with the negative control(NC)group,overexpression of miR-17-5p inhibited the expression of MMP-2 and significantly inhibited the number of clone formation,invading cells and migrating cells(P<0.05).After miR-17-5p was knocked down,the results were reversed.Conclusion:The expression of miR-17-5p is decreased in EMs interstitial cells.Inhibition of miR-17-5p can promote the expression of MMP-2 and enhance cell proliferation,invasion,and migration ability.