This study was aimed to investigate the effects of miR-650 on the proliferation, migration, invasion and apoptosis of rheumatoid arthritis synovial fibroblasts (RASFs).
Synovial tissue samples were collected from 16 rheumatoid arthritis (RA) patients and 13 patients with joint trauma undergoing joint replacement surgery. The RASFs were isolated and cultured. MiR-650 and AKT2 expressions in both synovial tissues and cells were detected using the quantitative real-time PCR (qRT-PCR), western blot and immunohistochemistry (IHC). Dual luciferase reporter gene assay was employed to evaluate the effect of miR-650 on the luciferase activity of AKT2. Then RASFs were transfected with miR-650 mimics, miR-650 inhibitors and pLenti6/V5-AKT2 respectively. And changes in cellular proliferation, migration, invasion and apoptosis were detected through MTT assay, wound healing assay, Transwell invasion assay and flow cytometry analysis, respectively.
MiR-650 was significantly down-regulated in RASFs than normal cells, whereas AKT2 was up-regulated in RASFs. Dual luciferase reporter gene assay showed that miR-650 could specifically bind to the 3'UTR of AKT2 and significantly repress the luciferase activity. MiR-650 significantly decreased the expression of AKT2. Down-regulation of miR-650 or up-regulation of AKT2 could increase proliferation, migration, invasion of RASFs, and decrease RASFs apoptosis. The conversed results were observed, when miR-650 was up-regulated in RASFs.
MiR-650 could inhibit the proliferation, migration and invasion of RASFs through targeted regulation of AKT2 expression.