miR-27a-3p对子宫内膜异位症异位内膜细胞迁移、侵袭和上皮间充质转化的影响

Effects of miR-27a-3p on ectopic endometrial cell migration, invasion and epithelial mesenchymal transformation in endometriosis

目的探究miR-27a-3p对子宫内膜异位症异位内膜细胞迁移、侵袭和上皮间充质转化的影响及可能机制。方法收集子宫内膜异位症患者在位内膜及异位内膜组织,qRT-PCR检测组织miR-27a-3p和YAP1mRNA表达;生物信息学预测YAP1 mRNA与miR-27a-3p的潜在结合位点,双荧光素酶报告基因实验验证;将miR-27a-3p模拟物及YAP1过表达质粒转染原代培养的异位内膜组织子宫内膜基质细胞,qRT-PCR检测细胞miR-27a-3p和YAP1 mRNA表达,Western blot检测细胞YAP1蛋白表达,CCK-8检测细胞增殖活性,Transwell实验检测细胞迁移和侵袭,Western blot检测细胞EMT标志物以及TGF-β2/Smad2/3信号通路相关蛋白表达。结果miR-27a-3p在异位内膜组织中低表达,YAP1 mRNA在异位内膜组织中高表达;miR-27a-3p与YAP1 mRNA靶向结合;转染miR-27a-3p模拟物增加异位内膜组织子宫内膜基质细胞miR-27a-3p表达,降低细胞YAP1 mRNA和蛋白表达,抑制细胞增殖、迁移和侵袭,且降低细胞N-cadherin、Vimentin、TGF-β2、p-Smad2和p-Smad3蛋白表达,增加细胞E-cadherin蛋白表达;过表达YAP1增加转染miR-27a-3p模拟物的子宫内膜基质细胞YAP1 mRNA和蛋白表达,促进细胞增殖、迁移和侵袭,且增加细胞N-cadherin、Vimentin、TGF-β2、p-Smad2和p-Smad3蛋白表达,降低细胞E-cadherin蛋白表达。结论MiR-27a-3p抑制子宫内膜异位症患者异位内膜细胞增殖、迁移、侵袭和EMT,其机制可能与靶向抑制YAP1表达抑制TGF-β2/Smad2/3信号通路有关。

Objective To explore the effects and possible mechanisms of miR-27a-3p on ectopic endometrial cell migration,invasion and epithelial mesenchymal transformation in endometriosis.Methods Tissues of endometriosis patients in situ and ectopic endometriosis were collected,and the expressions of miR-27a-3p and YAP1 mRNA were detected by qRT-PCR.The potential binding sites of YAP1 mRNA and miR-27a-3p were predicted by bioinformatics,and double luciferase reporter gene assaywas used to verified the combination.miR-27a-3p mimics and YAP1 overexpression plasmid were transfected into primary cultured ectopic endometrial tissue endometrial stromal cells.The expressions of miR-27a-3p and YAP1 mRNA were detected by qRT-PCR.The expression of YAP1 was detected by Western blot.Proliferation of cells was detected by CCK-8.The migration and invasion of cells were detected by Transwell assay.The expressions of EMT and TGF-β2/Smad2/3 signaling pathway related proteins were detected by Western blot.Results The expressions of miR-27a-3p was decreased in the ectopic endometrial tissues,and the expression of YAP1 mRNA was increased.miR-27a-3p binds to YAP1 mRNA.Transfection of miR-27a-3p mimics increased the expression of miR-27a-3p,decreased the expression of YAP1 mRNA and protein,and inhibited cell proliferation,migration and invasion of endometrial stromal cells in ectopic endometrial tissue.The expressions of N-cadherin,Vimentin,TGF-β2,p-Smad2 and p-smad3 proteins were decreased,and the expression of E-cadherin was increased.Overexpression of YAP1 increased the expression of YAP1mRNA and protein,promoted cell proliferation,migration,and invasion,increased the expressions of N-cadherin,Vimentin,TGF-β2,p-Smad2,and p-Smad3,and decreased the expression of E-cadherin in endometrial stromal cells transfected with miR-27a-3p mimics.Conclusion endometrial cells in patients with endometriosis,and its mechanism may be related to the inhibition of TGF-β2/Smad2/3 signaling pathway by targeted inhibition of YAP1 expression.