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

Background: 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.

Methods: 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.

Results: 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.

Conclusions: 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.