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作 者:陈礼荣[1] 范秋玉 刘亚 杨惠琴[1] CHEN Lirong;FAN Qiuyu;LIU Ya;YANG Huiqin(Department of Rheumatology and Immunoloy,Wuhan No.1 Hospital,Wuhan 430022,China)
出 处:《中国免疫学杂志》2025年第3期600-604,共5页Chinese Journal of Immunology
摘 要:目的:探究抑制miR-203a-3p基因表达对类风湿关节炎(RA)滑膜成纤维细胞增殖与凋亡的影响。方法:将成纤维样滑膜细胞MH7A分为Control组(未转染细胞)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-203a-3p组(转染anti-miR-203a-3p)、anti-miR-203a-3p+LiCl组(转染anti-miR-203a-3p+信号通路激活剂LiCl)。qRT-PCR检测miR-203a-3p表达水平;克隆形成实验、MTT实验检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测PCNA、Bcl-2、Bax、Wnt1、β-catenin蛋白表达。结果:转染miR-203a-3p可降低MH7A细胞克隆形成数、存活率,增加细胞凋亡率,降低PCNA、Bcl-2、Wnt1、β-catenin蛋白表达水平,增加Bax蛋白表达水平。信号通路激活剂LiCl增加转染miR-203a-3p对MH7A细胞克隆形成数、存活率,降低细胞凋亡率,增加Wnt1、β-catenin、PCNA、Bcl-2蛋白表达水平,降低Bax蛋白表达水平。结论:miR-203a-3p可能通过激活Wnt/β-catenin促进RA滑膜成纤维细胞增殖和抑制细胞凋亡。Objective:To explore the effect of inhibiting miR-203a-3p gene expression on proliferation and apoptosis of rheumatoid arthritis(RA)synovial fibroblasts.Methods:Divided fibroblast-like synovial cells MH7A into Control group(untransfected cells),anti-miR-NC group(transfected with anti-miR-NC),anti-miR-203a-3p group(transfected with anti-miR-203a-3p),anti-miR-203a-3p+LiCl group(transfected with anti-miR-203a-3p+signal pathway activator LiCl).qRT-PCR was used to detect expression level of miR-203a-3p;clone formation experiment,MTT experiment were used to detect cell proliferation;flow cytometry was used to detect cell apoptosis;Western blot was used to detect protein expressions of PCNA,Bcl-2,Bax,Wnt1,β-catenin.Results:Transfection of miR-203a-3p reduced the number of clones and survival rate of MH7A cells,increased rate of apoptosis,reduced expression levels of PCNA,Bcl-2,Wnt1,β-catenin protein,increased expression level of Bax protein.The signaling pathway activator LiCl increases the number of clones and survival rate of MH7A cells transfected with miR-203a-3p,reduces the rate of apoptosis,increases the expression level of Wnt1,β-catenin,PCNA,Bcl-2 protein,and reduces expression level of Bax protein.Conclusion:miR-203a-3p may promote proliferation of RA synovial fibroblasts and inhibit cell apoptosis by activating Wnt/β-catenin.
关 键 词:miR-203a-3p 类风湿关节炎 增殖 凋亡
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