In vitro and in vivo transfection of melanoma cells B16-F10 mediated by cholesterol-based cationic liposomes.
In vitro and in vivo transgene expression in B16-F10 melanoma cells has been investigated using an original cationic liposome prepared with triethyl aminopropane carbamoyl cholesterol iodide (TEAPC-Chol) as carrier. TEAPC-Chol/DOPE (dioleoyl phosphatidyl ethanolamine) liposomes are unilamellar, very stable and not toxic in the used concentration range. The yield in complexation with plasmid DNA can reach 100% even in the presence of fetal calf serum. The transfection level has been evaluated by luminometric measurements of luciferase expression. With TEAPC-Chol/DOPE (1:1) liposomes, a relatively high transfection level in B16-F10 cells has been observed comparing to commercial reagents. For in vivo assays, the transfection level in tumors induced in Nude mice has been optimized by studying the effects of charge ratio, of the helper lipid and of the injection volume. Results showed that TEAPC-Chol/DOPE (1:1) liposomes have improved 10-fold transfection level versus direct gene transfer of free DNA.
Reynier
P
P
Laboratoire de Chimie Structurale et Spectroscopic Biomoléculaire, CNRS, FRE 2313, UFR de Médecine, Université Paris XIII, 74 rue Marcel Cachin, F93017 Bobigny Cedex, France.
Briane
D
D
Cao
A
A
Lievre
N
N
Naejus
R
R
Bissieres
P
P
Salzmann
J L
JL
Taillandier
E
E
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002784:Cholesterol; D002850:Chromatography, Gel; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D007278:Injections, Spinal; D008081:Liposomes; D008164:Luminescent Proteins / Q000378:metabolism; D008546:Melanoma, Experimental / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010957:Plasmids / Q000235:genetics; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection
D004273:DNA, Neoplasm; D008081:Liposomes; D008164:Luminescent Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D049452:Green Fluorescent Proteins; D002784:Cholesterol; C022616:thiazolyl blue