摘要
目的探究miR-132-3p对类风湿关节炎成纤维样滑膜细胞(RA-FLSs)增值和侵袭的影响及调控机制。方法运用qRT-PCR分析miR-132-3p和卷曲类蛋白受体4(FZD4)RNA表达。将miR-132-3p模拟物、模拟物阴性对照、pcDNA3.1(+)载体以及FZD4过表达载体转染RA-FLSs后分析细胞活力、增殖、侵袭以及Wnt/β-catenin通路相关蛋白表达。再运用双荧光素酶报告实验鉴定miR-132-3p和FZD4关系。结果miR-132-3p在类风湿关节炎患者滑膜组织中的表达低于在正常滑膜组织中的表达,且在RA-FLSs中的表达低于在正常纤维样滑膜细胞中的表达,差异均有统计学意义(P<0.05);miR-132-3p模拟物转染后能抑制WT-FZD43’UTR组的荧光素酶活性,差异有统计学意义(P<0.05),但miR-132-3p模拟物对WT-FZD43'UTR组的荧光素酶活性无影响,差异无统计学意义(P>0.05);miR-132-3p转染组的RA-FLSs活力、EdU阳性细胞数、侵袭细胞数和β-catenin、c-Myc和CyclinD1蛋白表达明显低于miR-NC转染组,差异均有统计学意义(P<0.05),但是这些指标在miR-132-3p和FZD4共转染组中的表达又明显高于miR-132-3p和pcDNA共转染组,差异均有统计学意义(P<0.05)。结论MiR-132-3p通过抑制FZD4/Wnt/β-catenin通路激活来调控RA-FLSs增殖和侵袭。
Objective To investigate the effects of miR-132-3p on rheumatoid arthritis fibroblast-like synoviocyte(RA-FLSs)proliferation and invasion and the regulatory mechanism.Methods RNA expression of miR-132-3p and frizzled class receptor 4(FZD4)was analyzed by qRT-PCR.Cell viability,proliferation,invasion,and Wnt/β-catenin pathway-related protein expression were analyzed after transfection with miR-132-3p mimics,mimic negative control,pcDNA3.1(+)vector,and FZD4 overexpression vector.In addition,the relationship between miR-132-3p and FZD4 was identified by a dual-luciferase reporter assay.Results Compared with control group(synovium tissues or normal fibrous synovium cells),miR-132-3p was lowly expressed in synovial tissues and RA-FLSs of patients with rheumatoid arthritis,with statistically significant differences(P<0.05).miR-132-3p mimics significantly inhibited the luciferase activity of WT-FZD43'UTR group after transfection,with a statistically significant difference(P<0.05).However,miR-132-3p mimics had no effect on the luciferase activity of WT-FZD43'UTR group,and the difference was not statistically significant(P>0.05).The activity of RA-FLSs,the number of EdU positive cells,the number of invasive cells and the expression ofβ-catenin,c-Myc and CyclinD1 protein in miR-132-3p transfection group were significantly lower than those in miR-NC transfection group,with statistically significant differences(P<0.05).However,these indexes in the miR-132-3p and FZD4 co-transfection group were higher than those in the miR-132-3p and pcDNA co-transfection group,with statistically significant differences(P<0.05).Conclusion MiR-132-3p regulated the proliferation and invasion of RA-FLSs by inhibiting the activation of FZD4/Wnt/β-catenin pathway.
作者
梁舒
崔玉荣
吴洁
张超
郭占非
高晓
许振丹
范文强
LIANG Shu;CUI Yu-rong;WU Jie;ZHANG Chao;GUO Zhan-fei;GAO Xiao;XU Zhen-dan;FAN Wen-qiang(Department of Rheumatology and Immunology,Xinxiang Central Hospital&the Fourth Clinical College of Xinxiang Medical College,Xinxiang 453000,Henan,CHINA)
出处
《海南医学》
CAS
2023年第19期2743-2748,共6页
Hainan Medical Journal
基金
河南医学科技攻关计划(联合共建项目)(编号:LHGJ20210908、LHGJ20210904、LHGJ20210896)。
