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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 34, Number 3
BKCSDE 34(3)
March 20, 2013 

Stimulation of Phospholipase D in HepG2 Cells After Transfection Using Cationic Liposomes
Sang Yoon Lee*, Yan Lee, Joon Sig Choi, Jong Sang Park, Myung-Un Choi
Phospholipase D, Cationic liposomes, Cholesterol, Membrane perturbation
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.
931 - 935
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