Mechanism of microRNA-19a promoting the proliferation of osteoarthritis chondrocytes through first apoptosis signal/first apoptosis signal ligand signaling pathway
Objective To investigate the effects of microRNA-19a(miR-19a)on the proliferation and apoptosis of osteoarthritis(OA)chondrocytes by regulating the first apoptosis signal/first apoptosis signal ligand(Fas/FasL)pathway.Methods Chondrocytes were isolated from specimens of total knee arthroplas-ty,and both normal human chondrocytes and osteoarthritis(OA)chondrocytes were cultured in vitro.The expression of miR-19a was detected by real-time fluorescent quantitative polymerase chain reaction(RT-qPCR).OA chondrocytes were divided into control group,miR-19a negative control(NC)group,miR-19a mimic(mimics)group,miR-19a mimics+empty plasmid(pcDNA)group and miR-19a mimics+Fas overexpression plasmid(Fas)group.RT-qPCR method was used to detect the expression of miR-19a,Fas,FasL mRNA in each group of cells.Thiazole blue(MTT)method was used to detect the proliferation of OA chondrocytes in each group.Flow cytometry was used to detect cell apoptosis.Western blotting was used to detect the expression of cell proliferation,apoptosis-related proteins and Fas,FasL protein in each group.The dual luciferase reporter gene experiment was used to verify the targeting relationship between miR-19a and Fas.Data were analyzed using SPSS 22.0 software,and inter-group comparisons were con-ducted using t-tests and one-way analysis of variance(ANOVA),with a significance level set at P＜0.05.Results The expression level of miR-19a in the OA chondrocyte group was significantly lower than in the normal chondrocyte group(0.45±0.06 vs.1.01±0.19,t=2.95,P＜0.05).In the miR-19a mimics group,the expression levels of Fas and FasL were significantly lower than in the miR-19a NC group(Fas:0.31±0.05 vs.0.93±0.09,t=15.78,P＜0.05;FasL:0.37±0.04 vs.0.87±0.10,t=11.43,P＜0.05).The cell proliferation in the miR-19a mimics group was significantly higher than in the miR-19a NC group(A value:0.66±0.07 vs.0.45±0.05,t=8.70,P＜0.05),and the apoptosis rate in the miR-19a mimics group was significantly lower than in the miR-19a NC group[(8.57±1.65)％vs.(22.45±2.66)％,t=12.02,P＜0.05].Conclusion MiR-19a can promote the proliferation of OA chondrocytes and inhibit cell apoptosis by targeting the inhibition of Fas/FasL signaling pathway.
MicroRNAFirst apoptosis signal/first apoptosis signal ligand pathwayOsteoar-thritisChondrocytesProliferationApoptosis
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