肝靶向性纳米载体介导psiRNA对HepG2细胞HLA-E干扰效率的检测

Interfering HLA-E expression in HepG2 cell using psiRNA mediated by hepatocyte-targeted nano gene carrier

目的:检测肝细胞靶向性Gal-PEG-PEI纳米基因载体介导psiRNA干扰HepG2细胞HLA-E表达的效率。方法:分别用Gal-PEG-PEI/psiRNA、裸psiRNA、PEG-PEI/psiRNA、Gal-PEG-PEI/NC-psiRNA、Lipofectamine 2000/psiRNA转染HepG2细胞,同时以未转染的HepG2细胞为空白对照组,以Gal-PEG/NC-psiRNA(无义siRNA质粒)转染为阴性对照组。48 h后,Real-time RT-PCR检测HLA-EmRNA表达水平,Western blot法检测HLA-E蛋白表达水平。结果:Gal-PEG-PEI组对HLA-E的干扰效率为55%(mRNA水平)和62%(蛋白水平),显著高于Lipofectamine 2000组、PEG-PEI组、裸psiRNA组和阴性对照组(P<0.01)。结论:Gal-PEG-PEI/psiRNA纳米粒具有良好的肝细胞靶向性,对HepG2细胞HLA-E有较高的干扰效率。

Objective： To evaluate interference efficiency of galactosylated poly（ethylene glycol）-graft-polyethylenimine（Gal-PEG-PEI） as a nano gene carrier of hepatocyte-targeting plasmid siRNA（psiRNA） in HepG2 cells.Methods： HepG2 cells were transfected by Gal-PEG-PEI/psiRNA,naked-psiRNA,PEG-PEI/psiRNA,Gal-PEG-PEI/NC-psiRNA and Lipofectamine 2000/psiRNA respectively.The expression levels of HLA-E mRNA were detected by real time RT-PCR and HLA-E protein expression levels were detected by Western blot after 48 h.Results： The interference efficiencies of psiRNA mediated by Gal-PEG-PEI targetting HLA-E were 55%（mRNA expression level） and 62%（protein expression level）.They were distinctively higher than those of Lipofectamine2000,PEG-PEI and naked-psiRNA（P0.01）.Conclusion： The Gal-PEG-PEI/psiRNA nanospheres display perfect hepatocyte-targeting ability and have higher interference efficiency to HLA-E gene in HepG2 cells.