摘要
目的制备聚乙烯亚胺-三聚磷酸钠(PEI-TPP)纳米粒作为基因载体,并优化条件,获得高效转载表达质粒GFP-LC3(自噬标志性蛋白)的新型纳米粒基因载体。方法以阴离子凝聚交联法制备PEI-TPP纳米粒,通过静电吸附作用连接上质粒GFP-LC3;经琼脂糖凝胶电泳分析载体与DNA结合能力,并将其转染到常见细胞系观察其转染效率及细胞毒性。结果成功制备均匀分散、平均粒径为100 nm的PEI-TPP纳米粒。琼脂糖凝胶电泳的结果显示PEI-TPP纳米粒能有效地结合GFP-LC3。PEI-TPP纳米载体可以有效地转运LC3质粒进入Hela细胞并稳定表达。在常见细胞系中PEI-TPP纳米粒转染率比单一PEI转染高1.6~4.3倍,且细胞存活率达90%以上。结论 PEI-TPP纳米粒可有效结合并转染GFP-LC3质粒,呈现低毒和高效性,对研究细胞自噬的转染提供新的方法。
Objective To prepare and optimize the polyethyleneimine-sodium tripolyphosphate （PEI-TPP） nanoparticles as a new type gene carrier loading GFP-LC3 plasmid. Methods PEI was linked with TPP in anionic cross-linking way. The size, zata potential and other features of the particles were analyzed. And then the newly made nanoparticles were connected with plasmid GFP-LC3 by electrostatic adsorption. The combining ability with DNA of PEI-TPP nanopaticles was observed by agarose gel electrophoresis. In vitro cell transfection efficiency and cytotoxicity were also analyszed in Hela, 293T, HepG2 and MCF- 7 cells. Results Uniform dispersed PEI- TPP nanopaticles were successfully prepared, with average size of 100 nm. The electrophoresis results showed that PEI-TPP particles could effectively combined with GFP-LC3 plasmids and transfer the plasmids into Hela cells. Furthermore, in vitro test, the cellular transfection rate of PEI-TPP were 1.6-4.3 times higher than single PEI and with more than 90% cell survival rate. Conclusion The newly prepared PEI-TPP nanoparticles showed excellent characteristics as a new type of nanometer gene carrier, which may advance the study of autophagy.
作者
沈素晶
沈素云
李霭文
黎翠翠
庄丽颖
SHEN Su-jing;SHEN Su-yun;LI Ai-wenI;LI Cui-cui;ZHUANG Li-ying(I.Guangdong Second Trandiction Hospital,Guangzhou,Guangdong 510095;School of Engineering,Sun Yet-sen University,Guangzhou,Guangdong 510006;People＇s Hospital of Longhua,Shenzhen,Guangdong 518000,China)
出处
《热带医学杂志》
CAS
2018年第9期1147-1150,F0003,共5页
Journal of Tropical Medicine
