慢病毒介导shRNA沉默人纤维介素基因对心肌微血管内皮细胞增殖、迁移的影响

Effect of shRNA silencing fgl2 gene induced by lentivirus on microvascular endothelial cells of myocardial proliferation and migration

目的探讨人纤维介素(fgl2)对心肌微血管内皮细胞(MMVECs)增殖、迁移的影响。方法制作并包装fgl2的RNA干扰慢病毒,转染MMVECs。实验分为单纯MMVECs(对照组)、空载质粒慢病毒(GFP组)、fgl2短发夹RNA(shRNA)慢病毒(fgl2-RNAi-LV组)。稳定转染4 d后,实时定量RT-PCR(qRT-PCR)检测各组fgl2 mRNA表达,MTT检测细胞增殖;Transwell检测细胞迁移能力。结果 qRT-PCR结果显示,与对照组与GFP组比较,fgl2-RNAi-LV组的fgl2表达明显下调,MMVECs的增殖能力与迁移能力增强(P<0.05),而对照组与GFP组无明显差异。结论转染fgl2shRNA慢病毒载体后MMVECs的fgl2表达水平明显下调,fgl2shRNA慢病毒转染MMVECs,MMVECs的增殖能力与迁移能力增强。

Objective To construct RNAi lentivirus vector of fgl 2 gene and explore the effect of fgl 2 gene on microvascular endothe-lial cells ( MMVECs) of myocardial proliferation and migration.Methods RNAi lentivirus vector of fgl 2 gene was constructed and infected into MMVECs.Cells were divided into pure MMVECs ( control group ) , empty plasmid lentivirus ( GFP group ) , fgl2 short hairpin RNA (shRNA group), lentivirus (fgl2-RNAi-LV group).4 days later transfected into cells, quantitative real-time RT-PCR (Qrt-PCR) was used to detect fgl2 mRNA expression , MTT was used to detect cell proliferation , Transwell was used to detect cell migration.Results Compared with control and GFP groups , fgl2 expression was down-regulated and proliferation and migration of MMVECs were increased in fgl 2-RNAi-LV group ( P<0.05 ).There were no significant differences in above indexes between control and GFP groups .Conclusions fgl2 expression in MMVECs transfected with fgl 2-RNAi-LV is down-regulated and proliferation and migration of MMVECs are increased.