PEI与脂质体介导基因转染的比较研究

Comparison of gene transfection reagents between PEI and lipofectamine

目的比较新型转染试剂阳离子聚合物聚乙烯亚胺(PEI)与脂质体2000介导的血管内皮细胞生长因子VEGF165基因转染人胚肾细胞293T的转染效率,为基因转染提供新的方法。方法分别采用脂质体和不同N/P比值的PEI将合成的含有绿色荧光蛋白(GFP)和VEGF165的质粒转入293T细胞。转染24h后,采用CCK-8试剂盒检测细胞活性;转染48h后在荧光显微镜下观察比较两种转染方法的转染效率,并通过ELISA方法检测细胞上清液中VEGF165浓度,比较两种方法的转染效果。结果当N/P=9时,PEI转染的细胞活性与脂质体转染相近,对细胞的毒性较小;此时,PEI转染效率与脂质体也相似,均可达到约70%;同样,转染后细胞上清中VEGF165浓度较未转染组提高(P<0.05),但两种方法转染组间比较差异无统计学意义(P>0.05)。结论 PEI作为一种新型的转染试剂,与脂质体相比,在一定比例范围内,具有转染效率高,对细胞毒性小且价格低廉的优势,有望成为基因转染的有效载体。

Objective To compare the transfection efficiency of cationic polyethylenimine（PEI）with Lipofectamine 2000 TM by using the plasmid DNA encoding vascular endothelial cell growth factor（VEGF165）gene in human embryonic kidney cell line293 T.Methods PEI of different N/P ratio and Lipofectamine 2000 TM were used to deliver the vector containing VEGF165to293 Tcells,respectively.Green fluorescent protein（GFP）gene was inserted into the vector as a report gene.Evaluation of cytoactive was performed by CCK-8assay 24 hafter transfection.The cells were observed by fluorescent microscope and the presence of VEGF165 in cell supernatant was detected by ELISA 48 hafter transfection.The transfection efficiency was calculated and compared.Results Similar cytoactive and best transfection efficiency could be obtained when N/P ratio was 9,the transfection efficiency was around 70%.Furthermore,the presence of VEGF165 increased significantly after transfection（P〈0.05）,but there was no significant difference between the two groups in which different transfection methods were adopted.Conclusion PEI as a novel oligofectamine reagent could mediate more efficient transfection compared with lipofectamine.It also has low cell-toxicity and low price and could be an ideal vector in gene delivery technology.