Effect of lncRNA MINCR on the proliferation, invasion and migration of rheumatoid arthritis synovial fibroblasts by targeting miR-584-3p / 中华风湿病学杂志

ABSTRACT

Objective:To study the effects of MYC-induced long non-coding RNA (MINCR) targeting miR-584-3p on the proliferation, invasion and migration of rheumatoid arthritis synovial fibroblasts (RASFs).Methods:Synovial tissue samples were collected from 25 rheumatoid arthritis (RA) patients and 25 patients with joint trauma undergoing joint replacement surgery. Expression of MINCR and miR-584-3p in synovial tissue was detected using Real-time quantitative polymerase chain reaction (PCR). RASFs were separated for in- vitro culture, and RASFs were transfected with MINCR small interfering RNA (si-MINCR), miR-584-3p mimics, si-MINCR and miR-584-3p inhibitors (anti-miR-584-3p). Changes in cellular viability, clone formation, migration and invasion were detected by cell counting kit (CCK-8), plate cloning experiment, scratch healing test, Transwell test, respectively. Dual luciferase reporter gene assay was applied to evaluate the effect of miR-584-3p on the luciferase activity of MINCR. The independent t-test was used to analyze the differences between the two groups, and the one-way analysis of variance (ANOVA) and SNK- q test were used to analyze the differences between multiple groups. Results:MINCR was significantly up-regulated in RA synovial tissue compared to normal synovial tissue [(3.27±0.36) vs (1.00±0.08), t=30.777, P<0.01], whereas miR-584-3p was significantly down-regulated in RA synovial tissue [(0.43±0.05) vs (1.00±0.06), t=36.491, P<0.01]. After interference with MINCR expression, the activity [(0.52±0.04) vs (1.05±0.09), t=16.144, P<0.01] and clone formation number [(45±5) vs (99±9), t=15.960, P<0.01], scratch healing rate [(28±3)% vs (69±6)%, t=18.013, P<0.01] and invasion number [(53±5) vs (113±12), t=14.019, P<0.01] of RASFs were significantly decreased. After overexpression of miR-584-3p, the activity [(0.65±0.05) vs (1.02±0.09), t=26.063, P<0.01], clone formation number [(52±5) vs (98±8), t=14.619, P<0.01], scratch healing rate [(35±3)% vs (68±6)%, t=13.711, P<0.01] and invasion number [(62±5) vs (117±11), t=13.346, P<0.01] of RASFs were significantly decreased. miR-584-3p could specifically bind to MINCR and significantly inhibit the luciferase activity [(0.45±0.04) vs (0.97±0.06), t=21.633, P<0.01]. Compared with interference with MINCR, the activity [(0.92±0.07) vs (0.49±0.04), t=16.000, P<0.01], the clone formation number [(86±8) vs (45±5), t=14.008, P<0.01], scratch healing rate [(59±7)% vs (27±3)%, t=13.200, P<0.01], and invasions number [(99±9)% vs (54±5)%, t=13.414, P<0.01] of RASFs were significantly increased after miR-584-3p and MINCR were inhibited simultaneously. Conclusion:Interfering lncRNA MINCR can inhibit the proliferation, migration and invasion of RASFs by targeting and up-regulating miR-584-3p.