miR-27a-3p介导类风湿关节炎滑膜成纤维细胞增殖和侵袭的分子机制研究  

Study on the molecular mechanism of miR-27a-3p mediating proliferation and invasion of synovial fibroblasts in rheumatoid arthritis

作　　者：李宁宁[1] 雷蕾[1] 郝冬林[2] 刘喆 戴冰冰[1] LI Ningning;LEI Lei;HAO Donglin;LIU Zhe;DAI Bingbing(Department of Rheumatology and Immunology,Dalian Central Hospital,Dalian,Liaoning 116083,China;Department of Rheumatology and Immunology,Suzhou Traditional Chinese Medicine Hospital,Suzhou,Jiangsu 215008,China;Department of Rheumatology and Immunology,Zhumadian Central Hospital,Zhumadian,Henan 463000,China)

机构地区：[1]大连市中心医院风湿免疫科,辽宁大连116083 [2]苏州市中医医院风湿免疫科,江苏苏州215008 [3]驻马店市中心医院风湿免疫科,河南驻马店463000

出　　处：《重庆医学》2022年第4期546-550,共5页Chongqing medicine

基　　金：江苏省中医药管理局课题(YB2020057)。

摘　　要：目的探讨微RNA-27a-3p(miR-27a-3p)调控类风湿关节炎滑膜成纤维细胞(RASFs)增殖、侵袭的分子机制。方法体外分离培养正常滑膜成纤维细胞(SFs)和RASFs。实时荧光定量PCR(RT-PCR)检测miR-27a-3p表达;预测miR-27a-3p可能结合的潜在靶基因,并通过荧光素酶报告试验进行验证;通过Lipofectamine 2000将miR-27a-3p抑制剂转染入RASFs;分别采用细胞计数试剂盒8(CCK-8)、Transwell检测miR-27a-3p和靶基因对RASFs增殖、侵袭的影响。结果与SFs比较,miR-27a-3p在RASFs中过表达,miR-27a-3p可通过作用于分泌型卷曲相关蛋白1(SFRP1)的3′非翻译区抑制其表达;SFRP1在RASFs中弱表达,抑制miR-27a-3p表达后,SFRP1表达增加,细胞增殖和侵袭能力明显减弱(P<0.05)。同时抑制SFRP1后,增殖和侵袭能力明显恢复(P<0.05)。结论miR-27a-3p可促进RASFs的增殖与侵袭能力,对类风湿关节炎的发生起着重要作用,其机制可能是通过抑制SFRP1的表达实现的。Objective To investigate the molecular mechanisms of microRNA-27a-3p(miR-27a-3p)regulating the proliferation and invasion of rheumatoid arthritis synovial fibroblasts(RASFs).Methods Normal synovial fibroblasts(SFs)and RASFs were isolated and cultured in vitro.MiR-27a-3p expression was detected by real-time fluorescence quantitative PCR(RT-PCR).Potential target genes which miR-27a-3p might bind to were predicted and validated by luciferase reporter assay.The miR-27a-3p inhibitor was transfected into RASFs by Lipofectamine 2000.The effects of miR-27a-3p and target genes on the proliferation and invasion of RASFs were examined by Cell Counting Kit 8(CCK-8)and Transwell,respectively.Results Compared with SFs,miR-27a-3p was overexpressed in RASFs,and miR-27a-3p inhibited the expression of secretory frizzled-related protein 1(SFRP1)by acting on the 3′untranslated region of SFRP1.SFRP1 was weakly expressed in RASFs.After inhibition of miR-27a-3p expression,the SFRP1 expression was increased,and the cell proliferation and invasive ability was significantly reduced(P<0.05).The proliferative and invasive ability was significantly restored after simultaneous inhibition of SFRP1(P<0.05).Conclusion MiR-27a-3p can promote the proliferation and invasive ability of RASFs and play an important role in the development of rheumatoid arthritis,which probably through the inhibition of SFRP1 expression.

关 键 词：类风湿关节炎 微RNA-27a-3p 滑膜成纤维细胞 增殖 侵袭 分泌型卷曲相关蛋白1 

分 类 号：R593.22[医药卫生—内科学]