文章摘要

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

作者: 1洪哲晶, 1汪玲莉, 1郑纾
1 福建省人民医院妇科,福州 350004
通讯: 汪玲莉 Email: gxtm9a@163.com
DOI: 10.3978/j.issn.2095-6959.2021.03.001

摘要

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,Transwell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。
关键词: 子宫内膜异位症;miR-17-5p;MMP-2;子宫内膜异位症间质细胞;增殖;侵袭;迁移

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

Authors: 1HONG Zhejing, 1WANG Lingli, 1ZHENG Shu
1 Department of Gynecology, Fujian Provincial People’s Hospital, Fuzhou 350004, China

CorrespondingAuthor: WANG Lingli Email: gxtm9a@163.com

Abstract

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.
Keywords: endometriosis; miR-17-5p; MMP-2; endometriosis stromal cells; proliferation; invasion; migration