聚乙烯亚胺促进腺病毒体外基因表达

Polyethylenimine Polymer Promotes Gene Transduction of Adenovirus in Vitro

目的筛选较合适相对分子质量的聚乙烯亚胺(PEI)用于提高腺病毒载体基因表达效率。方法首先以物理吸附法使PEI与腺病毒相吸附,用噻唑蓝(MTT)法考察PEI-600、PEI-1 800、PEI-25 000复合腺病毒对细胞的毒性;利用体外A549细胞转导实验确定PEI-600、PEI-1 800和PEI-25 000复合腺病毒的最佳剂量比,并对复合物的Zeta电位和粒径分布进行考察分析。结果 MTT实验证明PEI相对分子质量越大,细胞毒性越大,其中PEI-25 000毒性最大;细胞转导试验中,PEI-1 800相对于PEI-600和PEI-25 000更显著促进腺病毒基因在A549细胞中的表达;Zeta电位测定结果显示PEI可以将腺病毒的表面负电荷转变为表面正电荷的粒子。结论在3种聚合物/腺病毒的复合物中,细胞毒性与聚合物的相对分子质量和浓度呈正相关。相对分子质量越大,浓度越大则聚合物对细胞毒性越大。

Objective To select polyethylenimine polymer with proper molecular weight to enhance the gene expression efficiency of Adenovirus （ Ad）. Methods Ad was resepcitively complexed with PEI-600,PEI-1 800 and PEI 25 000 by electrostatic adsorption. After^-ards, MTT （3-[4,5-dimethylthiazol-2-yl ] -2,5-diphenyltetrazolium bromide） assay was used to investigate the cytotoxicity of the complexes with a serial concentration, follow-ed by cell transduction assay to select the best amount of PEI with fixed dose of Ad.In the further,the physicochemical characteristics of the optimized complexes,Zeta potential, and size distribution were studied. Results MTT showed the higher the PEI moderate molecular weight was, the greater the cytotoxity was.PEI-25 000 showed a greater cytotoxicity than that of PEI-600 and PEI-1 800.As compared with PEI-600 and PEI- 25 000,PEI-1 800 significanlty promoted expression of Ad gene in A549 cells.PEI turned the Zeta potential of Ad from negative to positive charge. Conclusion In the three kinds of polymer/adenovirus compounds, the cytotoxicity of the compound is positively correlated with the polymer molecular weight and concentration. The greater molecular weight the polymer as well as the higher the concentration are,the higher the cytotoxicity is.