Stimulation of Phospholipase D in HepG2 Cells After Transfection Using Cationic Liposomes

Bulletin of the Korean chemical society

Myung-Un Choi¹  Sang Yoon Lee¹  Joon Sig Choi¹  Yan Lee¹  Jong Sang Park¹ 

关键词: Phospholipase D;    Cationic liposomes;    Cholesterol;    Membrane perturbation;   

DOI  :  

学科分类：化学（综合）

来源: Korean Chemical Society

【 摘 要 】

Lipid events in liposome-mediated transfection (lipofection) are largely unknown. Here we studied whether phospholipase D (PLD), an important enzyme responsible for phospholipid breakdown, was affected during lipofection of HepG2 cells with a luciferase plasmid. Synthetic cholesterol (Chol) derivatives, including 3β[Lornithinamide- carbamoyl]Chol, [polyamidoamine-carbamoyl]Chol and 3β[N-(N`,N`-dimethylaminoethane)- carbamoyl]Chol, and a cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride were mixed with a helper lipid dioleoylphosphatidylethanolamine to form respective cationic liposomes. All cationic liposomes were found to stimulate PLD. Although orders of magnitude effects of the cationic liposomes on PLD stimulation did not consistently match those on cytotoxicity and luciferase expression, a causal relationship between PLD activation and cytotoxic effect was remarkable. PLD stimulation by the cationic liposomes was likely due to their amphiphilic characters, leading to membrane perturbation, as supported by similar results obtained with other membrane-perturbing chemicals such as oleate, melittin, and digitonin. Our results suggest that lipofection induces cellular lipid changes such as a PLD-driven phospholipid turnover.

【 授权许可】

Unknown