PS exposure increases the susceptibility of cells to fusion with DOTAP liposomes.
Cationic liposomes are used as efficient carriers for gene delivery into mammalian cells due to their ability to bind nucleic acids, adsorb onto the cell surface and fuse with negatively charged membranes. This last property enables the release and escape of their cargo from endosomal compartments. The efficiency of this fusion mainly depends on the surface charge of the target membranes. Here, we report that cells of two different lines, epithelial adenocarcinoma HeLa and lymphocytic leukemia Jurkat T, which externalize PS, are more susceptible to fusion with DOTAP liposomes than control cells. We compared the ability to undergo fusion of untreated and apoptotic cells. Apoptosis was induced by various pro-apoptotic agents and treatments, namely: incubation in the presence of MnCl(2), cytostatic drugs fludarabine and mitoxantrone, staurosporine and serum depletion in the case of HeLa cells. Jurkat T cells were treated similarly except apoptosis was additionally induced by incubation in the presence of 4% EtOH. Epithelial cells fused with the highest efficiencies of lipid mixing, when pretreated with staurosporine. Jurkat T cells were less susceptible to fusion, but they also displayed an increase in fusion efficiency after the induction of apoptosis. Alternatively, we treated the cells with metabolic inhibitors causing ATP-depletion in order to inactivate aminophospholipid translocase. After ATP-depletion, HeLa and Jurkat T cells fused with DOTAP liposomes with higher efficiencies than control cells. Our conclusion is that the lipid asymmetry of natural membranes may limit fusion with cationic liposomes.
Laboratory of Cytobiochemistry, Institute of Biochemistry and Molecular Biology, University of Wroclaw, Przybyszewskiego 63/77, 51-148 Wroclaw, Poland.
Katarzyna
K
Stebelska
Paulina
P
Wyrozumska
Aleksander F
AF
Sikorski
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects; D002712:Chlorides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D008081:Liposomes; D017895:Manganese Compounds / Q000494:pharmacology; D008561:Membrane Fusion; D008942:Mitoxantrone / Q000494:pharmacology; D010718:Phosphatidylserines / Q000737:chemistry* / Q000378:metabolism; D049294:Phospholipid Transfer Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000737:chemistry* / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology; D014740:Vidarabine / Q000031:analogs & derivatives / Q000494:pharmacology
D000970:Antineoplastic Agents; D002712:Chlorides; D004791:Enzyme Inhibitors; D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D008081:Liposomes; D017895:Manganese Compounds; D010718:Phosphatidylserines; D049294:Phospholipid Transfer Proteins; D000644:Quaternary Ammonium Compounds; D008942:Mitoxantrone; D014740:Vidarabine; D019311:Staurosporine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; C024352:fludarabine; C025340:manganese chloride