miR-218-5p in Endometrial Microenvironment Prevents the Migration of Ectopic Endometrial Stromal Cells by Inhibiting LASP1

Abstract

Our previous two-dimensional electrophoresis experiments showed that the expression of LASP1 in patients with endometriosis was significantly higher than that of control endometrium. However, the molecular mechanism by which LASP1 is regulated in endometriosis/adenomyosis is unknown. Herein, qPCR was performed to analysis the expression levels of LASP1 and miR-218-5p between EMs cells and control cells. Immunofluorescence in situ hybridization was carried out to measure the expression level of miR-218-5p in ectopic endometrium versus normal endometrium. After miR-218-5p mimic or inhibitor were transfected, the transwell experiment was carried out to see the effect of miR-218-5p on the migration of ESCs. EdU was used to measure cell proliferation rate. Dual-luciferase reporter assay (DR) was used to verify the binding of hsa-miR-218-5p to the 3'UTR of LASP1. Western blot and immunofluorescence analysis were carried out to identify the protein expression pattern of LASP1 and EMT markers in endometrial tissue. Our results show that high expression of LASP1 is related to the development of EMs. miR-218-5p inhibits the expression level of LASP1 by binding the 3'UTR region of LASP1. miR-218-5p is negatively correlated with the expression of LASP1 protein in endometriosis cells and tissues. Without affecting proliferation, miR-218-5p can increase the migration of ESCs. miR-218-5p is mainly secreted from blood vessels and expressed in the muscle layer around the endometrium. miR-218 also inhibits mesenchymal transition of epithelial cells by inhibiting the expression of Vimentin.