Downregulating miR-96-5p promotes proliferation, migration, and invasion, and inhibits apoptosis in human trophoblast cells via targeting DDAH1

Abstract

Several microRNAs (miRs) have been found to have modulating effects on trophoblast functions, yet the biological role and function of miR-96-5p and its interaction with Dimethylarginine Dimethylaminohydrolase 1 (DDAH1) remained poorly understood.After lentivirus transfection, the proliferation, migration, invasion and apoptosis of human trophoblast cells HTR-8/SVneo and SGHPL-4 were determined by Cell Counting Kit-8 (CCK-8) assay, scratch assay, Transwell, and flow cytometry, respectively. Relative expressions of miR-96-5p, DDAH1, and apoptosis-related proteins (B-cell lymphoma 2, Bcl-2; Bcl-2-associated X protein, Bax; cleaved (C) caspase-3) were detected via quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. The target gene of miR-96-5p and their potential binding sites were predicted using TargetScan V7.2 and confirmed by dual-luciferase reporter assay.MiR-96-5p downregulation promoted proliferation, migration and invasion, suppressed apoptosis, and decreased miR-96-5p expression in trophoblast cells in vitro, while miR-96-5p upregulation had the opposite effects. DDAH1 was recognized as a target gene of miR-96-5p, and silencing DDAH1 reversed the effects of miR-96-5p downregulation on the proliferation, migration, invasion and apoptosis of trophoblast cells as well as the expressions of apoptosis-related proteins.MiR-96-5p downregulation promotes proliferation, migration, and invasion, and suppresses apoptosis in human trophoblast cells in vitro via targeting DDAH1, which provides evidence for the implication of miR-96-5p in the functional modulation of trophoblasts.