靶向性半乳糖化聚乙二醇-聚乙烯亚胺纳米载体介导siRNA质粒对肝癌细胞BEL-7402的转染效率检测

Detection of transfection efficiency of targeting galactosylated poly(ethylene glycol)-graft-polyethylenimine as a nanoparticle carrier mediatig psiRNA in hepatocellular carcinoma BEL-7402 cells

目的检测靶向性半乳糖化聚乙二醇-聚乙烯亚胺(Gal-PEG-PEI)纳米载体介导siRNA质粒(psiRNA)对肝癌细胞BEL-7402的转染效率。方法纳米粒径仪测定Gal-PEG-PEI/psiRNA纳米粒的粒径和zeta电位;以Gal-PEG-PEI/psiRNA为研究组,以聚乙二醇-聚乙烯亚胺(PEG-PEI)/psiRNA、Lipofectamine2000和裸psiRNA转染为对照组,转染BEL-7402细胞,48h后用流式细胞仪测定各组的转染效率。转染前加入1mg半乳糖观察各组的半乳糖竞争拮抗结果。结果 Gal-PEG-PEI/psiRNA复合物的粒径随氨基与磷酸基比例(N/P)的增大而减小,最小粒径为81.1nm;Gal-PEG-PEI载体的转染效率为(24.34±2.55)%,与Lipofectamine2000组的转染效率(22.88±1.22)%比较差别无统计学意义(P>0.05),但明显高于PEG-PEI的转染效率(12.91±0.36)%及裸psiRNA的转染效率(0.73±0.08)%,差别有统计学意义(P<0.01);加入竞争拮抗剂半乳糖后Gal-PEG-PEI介导的转染效率由(24.34±2.55)%下降至(5.85±1.83)%,差别有统计学意义(P<0.01)。结论 Gal-PEG-PEI/psiRNA纳米粒对肝癌细胞BEL-7402有较高的转染效率,具有良好的肝细胞靶向性。

Objective To evaluate transfection efficiency of galactosylated poly（ ethylene glycol）-graft-polyethylenimine （ Gal-PEG-PEI） as a nanoparticle carrier mediating hepatocyte-targeting plasmid siRNA（ psiRNA） in hepatocellular carcinoma cells line BEL-7402. Methods The grain size and zeta poten of the Gal-PEG-PEI /psiRNA nanoparticles were measured by dy-namic light scattering. Gal-PEG-PEI /psiRNA were experimental group,PEG-PEI /psiRNA,Lipofectamine 2000 and naked psiR-NA were control group. BEL-7402 cells were transfected the transfection efficiency in two groups were evaluated by flow cytometry after 48 hours. The rivalry consequence assay was carried out after adding 1 mg galactose into the transfection media. Results The results indicated that the particle sizes decreased as charge ratios of Gal-PEG-PEI to psiRNA increasing and a minimum val-ue was around 81. 1 nm. The transfection efficiency of Gal-PEG-PEI and Lipofectamine 2000 were （ 24. 34 ？？2. 55 ） % and （ 22.88 ？？.22） % respectively,the difference was not statistical significant（ P 0. 05） ,and the transfection efficiency of Gal-PEG-PEI and Lipofectamine 2000 higher than that of PEG-PEI （ 12. 91 ？？0. 36） % or naked psiRNA （ 0. 73 ？？0. 08） % in BEL-7402 cells. When galactose was added,the transfection efficiency of Gal-PEG-PEI was drastically decreased from （ 24. 34 ？？2. 55） % to （ 5. 85 ？？1. 83） %. There was significant difference（ P 0. 01）. Conclusion The Gal-PEG-PEI /psiRNA nanospheres display perfect hepatocyte-targeting ability and have good transfection efficiency in hepatocellular carcinoma cells line BEL-7402.