hsa_circ_0044235对类风湿关节炎滑膜成纤维细胞增殖和迁移的影响及机制  

Effect of hsa_circ_0044235 on proliferation and migration of rheumatoid arthritis synovial fibroblasts and its mechanism

作　　者：田晓静[1] 薛宏峰[1] 蒋辉[1] 姚雨叶 TIAN Xiaojing;XUE Hongfeng;JIANG Hui;YAO Yuye(Department of Laboratory Medicine,Changzhou TCM Hospital Affiliated to Nanjing TCM University,Changzhou 213003,China)

机构地区：[1]南京中医药大学附属常州市中医医院检验科,常州213003

出　　处：《中国免疫学杂志》2022年第22期2712-2717,共6页Chinese Journal of Immunology

摘　　要：目的:探讨hsa_circ_0044235对类风湿关节炎滑膜成纤维细胞增殖和迁移的影响及可能机制。方法:体外培养滑膜成纤维细胞MH7A,分为对照组(Con组)、LPS组、LPS+pcDNA组、LPS+pcDNA-hsa_circ_0044235组、LPS+pcDNAhsa_circ_0044235+miR-NC组和LPS+pcDNA-hsa_circ_0044235+miR-338-5p组,RT-qPCR检测各组细胞hsa_circ_0044235和miR-338-5p表达,克隆形成实验检测细胞增殖,流式细胞术检测细胞周期,划痕实验检测细胞迁移,Western blot检测细胞E-cadherin和N-cadherin蛋白表达。双荧光素酶报告基因实验验证hsa_circ_0044235与miR-338-5p的调控关系。结果:与Con组比较,LPS组MH7A细胞hsa_circ_0044235表达和E-cadherin蛋白表达降低(P<0.05),而miR-338-5p表达、细胞克隆形成数、划痕愈合率及N-cadherin蛋白表达升高(P<0.05),细胞周期G_(0)-G_(1)期缩短(P<0.05),S期延长(P<0.05)。与LPS+pcDNA组比较,LPS+pcDNA-hsa_circ_0044235组MH7A细胞克隆形成数、划痕愈合率及N-cadherin蛋白表达降低(P<0.05),细胞周期G_(0)-G_(1)期延长(P<0.05),S期缩短(P<0.05),E-cadherin蛋白表达升高(P<0.05)。与LPS+pcDNA-hsa_circ_0044235+miR-NC组比较,LPS+pcDNA-hsa_circ_0044235+miR-338-5p组MH7A细胞克隆形成数、划痕愈合率及N-cadherin蛋白表达升高(P<0.05),细胞周期G_(0)-G_(1)期缩短(P<0.05),S期延长(P<0.05),E-cadherin蛋白表达降低(P<0.05)。双荧光素酶报告基因实验结果显示,hsa_circ_0044235可靶向结合miR-338-5p。结论:过表达hsa_circ_0044235可能靶向抑制miR-338-5p表达降低LPS诱导的滑膜成纤维细胞增殖和迁移,hsa_circ_0044235/miR-338-5p轴可能为类风湿关节炎治疗提供了新靶点。Objective:To investigate effect of hsa_circ_0044235 on proliferation and migration of rheumatoid arthritis synovial fibroblasts and its possible mechanism.Methods:Synovial fibroblasts MH7A were cultured in vitro and divided into control group(Con group),LPS group,LPS+pcDNA group,LPS+pcDNA-hsa_circ_0044235 group,LPS+pcDNA-hsa_circ_ 0044235+miR-NC group and LPS+pcDNA-hsa_circ_0044235+miR-338-5p group. RT-qPCR was used to detect cell expressions of hsa_circ_0044235 and miR-338-5p in each group,clone formation test was used to detect cell proliferation,flow cytometry was used to detect cell cycle,scratch test was used to detect cell migration,and Western blot was used to detect protein expressions of E-cadherin and N-cadherin.Dual luciferase reporter gene experiment was used to verify regulatory relationship between hsa_circ_0044235 and miR-338-5p.Results:Compared with Con group,expressions of hsa_circ_0044235 and E-cadherin protein of MH7A cells in LPS group were decreased(P<0.05),while expression of miR-338-5p,number of clone cells,scratch healing rate and protein expression of N-cadherin were increased(P<0.05),G_(0)-G_(1)phase of cell cycle was shortened(P<0.05),and S phase was prolonged(P<0.05). Compared with LPS+pcDNA group,number of MH7A cell clones,scratch healing rate and protein expression of N-cadherin in LPS+pcDNA-hsa_circ_0044235 group were decreased(P<0.05),G_(0)-G_(1)phase of cell cycle was prolonged(P<0.05),S phase was shortened(P<0.05),and expression of E-cadherin protein was increased(P<0.05). Compared with LPS+pcDNA-hsa_circ_0044235+miR-NC group,number of MH7A cell clones,scratch healing rate and protein expression of N-cadherin in LPS+pcDNA-hsa_circ_0044235+miR-338-5p group were increased(P<0.05),G_(0)-G_(1)phase of cell cycle was shortened(P<0.05),S phase was prolonged(P<0.05),and expression of E-cadherin protein was decreased(P<0.05). Dual luciferase reporter gene experiment results showed that hsa_circ_0044235 could targeting regulate miR-338-5p.Conclusion:Over-expression of hsa_circ_0044235 may

关 键 词：类风湿关节炎 滑膜成纤维细胞 hsa_circ_0044235 miR-338-5p 细胞增殖 迁移 

分 类 号：R593.22[医药卫生—内科学]