miR-23b-3p靶向XIAP调控类风湿关节炎滑膜成纤维细胞增殖和凋亡的机制研究

目的:探讨微小RNA-23b-3p(miR-23b-3p)靶向X连锁凋亡抑制蛋白(XIAP)对类风湿关节炎滑膜成纤维细胞增殖和凋亡的影响及其作用机制。方法:应用RT-qPCR检测体外培养的RA滑膜成纤维细胞中miR-23b-3p和XIAP的表达水平。通过TargetScan预测分析miR-23b-3p与XIAP的靶向关系,并使用双萤光素酶报告基因实验进行验证。采用脂质体法将miR-23b-3p模拟物和抑制物转入细胞中,RT-qPCR法检测miR-23b-3p和XIAP表达水平的变化;CCK-8法和细胞流式术分别检测miR-23b-3p对细胞活力和细胞凋亡的影响;Western blot检测Ki67和Bcl-2蛋白表达量的变化。结果:类风湿关节炎滑膜成纤维细胞中miR-23b-3p表达显著下调,XIAP表达显著上调(P<0.05)。转染miR-23b-3p模拟物促使XIAP表达显著下调(P<0.05),并且显著抑制细胞活力,促进细胞凋亡(P<0.05),同时Ki67和Bcl-2显著下调(P<0.05);转染miR-23b-3p抑制物的效果与之相反。结论:miR-23b-3p通过靶向XIAP抑制类风湿关节炎滑膜成纤维细胞增殖,促进其凋亡。

Effect of miR-23b-3p on viability and apoptosis of rheumatoid arthritis synovial fibroblasts by targeting XIAP

AIM: To investigate the effect of microRNA-23 b-3 p(miR-23 b-3 p) on the viability and apoptosis of rheumatoid arthritis synovial fibroblasts by targeting X-linked inhibitor of apoptosis protein(XIAP). METHODS: The expression of miR-23 b-3 p and XIAP was detected by RT-qPCR. The TargetScan was used to predict the targeting regulatory relation between miR-23 b-3 p and XIAP, and then the regulatory relation was confirmed by dual-luciferase reporter assay. After the miR-23 b-3 p mimic and inhibitor were transfected into the cells, the expression of miR-23 b-3 p and XIAP was detect by RT-qPCR. The effect of miR-23 b-3 p on the viability and apoptosis was measured by CCK-8 assay and flow cytometry. The protein expression levels of Ki67 and Bcl-2 were determined by Western blot. RESULTS: The expression level of miR-23 b-3 p was down-regulated significantly(P<0.05), and XIAP was up-regulated significantly in rheumatoid arthritis synovial fibroblasts(P<0.05). The miR-23 b-3 p mimic significantly inhibited XIAP expression and the cell viability, promoted the apoptosis, and down-regulated the expression of Ki67 and Bcl-2(P<0.05). The effects of miR-23 b-3 p inhibitor were the opposite. CONCLUSION: miR-23 b-3 p inhibits the viability and promotes apoptosis of rheumatoid arthritis synovial fibroblasts by targeting XIAP.