分枝状聚乙烯亚胺对不同细胞系的转染效率和细胞毒性作用

Effect of branch polyethylenimine on transfection and cytotoxicity in different cell lines

目的探讨分支状聚乙烯亚胺(PEI)作为载体对不同细胞株的转染效率和细胞毒性。方法培养人类胚胎肾细胞293T、人上皮癌细胞Hela、小鼠骨髓间充质干细胞(BMSC)和大鼠肾上腺嗜铬细胞瘤细胞PC12。悬浮转染p EGFP质粒与PEI的复合物,利用荧光细胞计数法分析转染效率;悬浮转染p Luc(luciferase基因来自Renilla)质粒与PEI的复合物,利用分光光度计分析各细胞的荧光素酶活性;采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法分析PEI对293T、Hela、BMSC和PC12细胞的毒性,分析转染效率和细胞毒性之间的相关性。结果293T、Hela、BMSC和PC12细胞的转染效率分别为(92.1±4.5)%、(29.2±3.4)%、(21.5±2.1)%和(17.0±3.2)%。293T、Hela、BMSC和PC12细胞的荧光素酶活性分别是2.87×108、3.35×107、1.94×107和1.08×107RLU·mg protein-1。293T、Hela、BMSC和PC12细胞转染效率排序:293T>Hela>BMSC>PC12细胞。同时,MTT比色法结果表明细胞毒性排序:293T>Hela>BMSC>PC12细胞。转染效率与细胞毒性呈正相关(r=0.865,P<0.05)。结论 PEI对不同细胞株的转染效率和细胞毒性存在一定的正相关性。

Objective To explore the effect of branched polyethyleneimine （PEI） as a trager on transfection and cyto- toxicity in different cell lines. Methods Four cell lines including 293T, Hela, bone mesenchymal stem cells （BMSC） and PC12 were cultured. The cells in suspension were transfected with complexes of plasmid pEGFP and PEI, then the transfection efficiency was analyzed by positive green fluorescent protein cell counting; meanwhile, various cells were transfected with com- plexes of plasmid pLuc （encoding luciferase of Renilla） and PEI,and luciferase activity was assessed by the spectrophotome- ter. And 3-（4,5-Dimethyhhiazol-2-yl）-2,5-diphenyhetrazolium bromide （MTF） test was used to deternmine cytotoxicity of PEI on the four cell lines. The correlation of transfection and cytotoxicity was analysed. Results Transfection efficiency of 293T, Hela, BMSC and PC12 was （92.1± 4.5 ） %, （29.2 ±3.4） %, （21.5± 2.1 ） %, （ 17.0± 3.2） %, respectively. Lucifer- ase activity of 293T, Hela, BMSC and PC12 was 2.87 ×10^8,3.35 ×10^7 , 1.94×10^7 , 1.08×10^7 RLU · mg protein - 1, respec- tively. The results of positive green fluorescent protein cell counting method and luciferase activity analysis demonstrated that the order of transfection efficiency was 293T 〉 Hela 〉 BMSC 〉 PC12 ceils. Also, the order of cytotoxicity was 293T 〉 Hela 〉 BMSC 〉 PC12 cells. The transfection efficiency was positive related with eytotoxieity（r = 0. 865,P 〈 0.05）. Conclusion The data strongly indicate that transfection efficiency is positively related to the cytotoxicity of PEI in different cells.