Long non-coding RNA IGF2-AS promotes trophoblast cell proliferation, migration, and invasion by regulating miR-520g/N-cadherin axis.

Abstract

Recurrent miscarriage (RM) is a distressing reproductive issue worldwide. Dysfunction of trophoblasts can trigger numerous unfavorable pregnant outcomes such as RM, stillbirth, and fetal malformation. In this text, the roles and molecular basis of long non-coding RNA insulin growth factor 2 antisense (IGF2-AS) in the development of trophoblast cells were further investigated. IGF2-AS, microRNA-520g (miR-520g), and N-cadherin levels were measured by RT-qPCR assay. Cell viability, the number of colonies, cell apoptosis, migration, and invasion were measured by CCK-8 assay, colony formation assay, flow cytometry, transwell migration, and invasion assays, respectively. The relative proteins expression was detected by western blot. The interaction between miR-520g and IGF2-AS or N-cadherin was tested by bioinformatics prediction analysis, and confirmed by dual-luciferase reporter assay, RNA pull-down assay, and RNA immunoprecipitation (RIP) assay. Our data revealed that IGF2-AS and N-cadherin levels were notably decreased, and miR-520g was strikingly increased in the placentas from RM patients. IGF2-AS overexpression promoted cell proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), and hampered cell apoptosis in trophoblast cells, while IGF2-AS deletion exhibited opposite results. Moreover, miR-520g was a target gene of IGF2-AS and negatively regulated by IGF2-AS. MiR-520g inhibitor enhanced the proliferation, migration, and invasion capability of trophoblast cells, suppressed cell apoptosis, and promoted the EMT process. Moreover, the effects of IGF2-AS overexpression on trophoblast cells were reversed by miR-520g upregulation. These findings indicated that IGF2-AS facilitated trophoblast cell proliferation, migration, invasion, EMT, and suppressed cell apoptosis by regulating miR-520g/N-cadherin axis, providing potential biomarkers for RM.