miR-214-5p靶向SEMA4C基因调控类风湿关节炎滑膜成纤维细胞增殖和侵袭的分子机制  

Molecular mechanism of miR-214-5p regulating the proliferation and invasion of rheumatoid arthritis synovial fibroblasts through targeting SEMA4C gene

作　　者：董宝铁[1] 李泓[1] 高伟[1] 周振东[1] DONG Baotie;LI Hong;GAO Wei(Department of Osteology,The Fourth People’s Hospital of Shenyang City,Liaoning,Shengyang 110031,China)

机构地区：[1]辽宁省沈阳市第四人民医院骨一科,110031

出　　处：《河北医药》2021年第9期1307-1311,共5页Hebei Medical Journal

摘　　要：目的研究miR-214-5p对类风湿性关节炎滑膜成纤维细胞(RA-FLSs)增殖和侵袭的影响和潜在的分子机制。方法根据转染物将RA-FLSs细胞分为miR-NC组、miR-214-5p组、si-NC组、si-SEMA4C组、anti-miR-NC组、anti-miR-214-5p组、miR-214-5p+pcDNA-NC组和miR-214-5p+pcDNA-SEMA4C组。qRT-PCR检测RA-FLSs中SEMA4C mRNA和miR-214-5p的表达,Western Blot检测SEMA4C、CyclinD1、MMP-2和MMP-9蛋白表达水平,CCK8法和Transwell实验分别测定RA-FLSs细胞增殖率和侵袭能力,双荧光素酶报告系统验证miR-214-5p与SEMA4C间的调控关系。结果与control组比较,在RA-FLSs细胞中miR-214-5p的表达量显著下降(P<0.05),而SEMA4C mRNA和蛋白的表达量显著升高(P<0.05);与对照miR-NC和si-NC组比较,miR-214-5p组和si-SEMA4C组RA-FLSs细胞的增殖率和侵袭细胞数均下降,CyclinD1、MMP-2和MMP-9蛋白水平均降低,差异均有统计学意义(P<0.05);双荧光素酶报告系统结果显示,miR-214-5p靶向负调控SEMA4C蛋白表达;与miR-214-5p+pcDNA-NC组比较,miR-214-5p+pcDNA-SEMA4C组RA-FLSs细胞增殖率和侵袭细胞数均升高,CyclinD1、MMP-2和MMP-9蛋白水平均升高,差异均有统计学意义(P<0.05)。结论miR-214-5p通过靶向SEMA4C基因调控RA-FLSs细胞增殖和侵袭,miR-214-5p有望成为类风湿性关节炎分子诊断和治疗的靶点。Objective To investigate the effects of miR-214-5p on the proliferation and invasion of rheumatoid arthritis synovialfibroblasts(RA-FLSs)and the possible molecular mechanism.Methods RA-FLSs cells were divided into miR-NC group,miR-214-5p group,si-NC group,si-SEMA4C group,anti-miR-NC group,anti-miR-214-5p group,miR-214-5p+pcDNA-NC group and miR-214-5p+pcDNA-SEMA4C group according to transfectants.The qRT-PCR was used to detect the expression levels of SEMA4C mRNA and miR-214-5p in RA-FLSs.Western Blot was used to detect the expression levels of SEMA4C,CyclinD1,MMP-2 and MMP-9 proteins.Moreover CCK8 and Transwell assay were used to detect the proliferation and invasion ability of RA-FLSs.Dual-luciferase reporter assay system was performed to verify the regulating correlation between miR-214-5p and SEMA4C.Results As compared with those in control group,the expression levels of miR-214-5p in RA-FLSs cells were decreased significantly(P<0.05),however,the expression levels of SEMA4C mRNA and protein were significantly increased(P<0.05).As compared with those in control miR-NC and si-NC group,the proliferation rate and the number of invasive cells of RA-FLSs cells in miR-214-5p group and si-SEMA4C group were significantly decreased,and the protein levels of CyclinD1,MMP-2 and MMP-9 were significantly decreased(P<0.05).The dual luciferase reporting system showed that miR-214-5p targetedly and negatively regulated the expression of SEMA4C protein.As compared with those in miR-214-5p+pcDNA-NC group,the proliferation rate and invasive cell number of RA-FLSs cells in miR-214-5p+pcDNA-SEMA4C group were significantly increased,and the levels of CyclinD1,MMP-2 and MMP-9 proteins were significantly increased(P<0.05).Conclusion The miR-214-5p can regulate the proliferation and invasion of RA-FLSs cells by targeting SEMA4C gene,and miR-214-5p is expected to be the new target of molecular diagnosis and treatment of rheumatoid arthritis.

关 键 词：类风湿性关节炎 miR-214-5p SEMA4C 增殖 侵袭 

分 类 号：R539.22[医药卫生—内科学]