Liver-targeted gene transfer into a human hepatoblastoma cell line and in vivo by sterylglucoside-containing cationic liposomes.
We investigated the transfection efficiency of beta-sitosterol beta-D-glucoside (Sit-G)-containing liposome/DNA complex (Sit-G-liposome/DNA complex) for liver targeting. The Sit-G-liposome/DNA complex was composed of Tfx-20 reagent (Tfx), ie synthetic cationic lipid [N,N,N',N'-tetramethyl-N,N'-bis(2-hydroxyethyl)-2,3-di(oleoyloxy)-1,4-butanediammonium iodide] with L-dioleoylphosphatidylethanolamine (DOPE), 3 beta[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol (DC-Chol) and Sit-G with plasmid DNA. The in vitro studies were performed in HepG2 cells in serum-containing medium and the in vivo studies were carried out in the mice following intravenous injection. The Sit-G-liposome produced a Sit-G-liposome/DNA complex of relatively small size (100--250 nm). Transfection efficiency of the luciferase marker gene by Sit-G-liposome/DNA complex was increased in the presence of 10% serum in vitro, and was selectively high in the mouse liver reaching expression values up to an average of 14.9 pg luciferase/mg tissue protein, compared with Tfx/DNA complex, which showed approximately three-fold higher gene expression than Sit-G-liposome/DNA complex in vitro. High in vitro transfection efficiency by Sit-G-liposome/DNA complex seemed to be possible even with large lipid precipitates, whereas high in vivo activity seemed to be related to small and dispersed complexes. The interaction of liposome/DNA complexes with serum may be a key point to predict the in vivo efficiency of a liposome vector.
Hwang
S H
SH
Department of Pharmaceutics, Hoshi University, Shinagawa-ku, Tokyo, Japan.
Hayashi
K
K
Takayama
K
K
Maitani
Y
Y
D000818:Animals; D002784:Cholesterol / Q000031:analogs & derivatives; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D020404:Glycerophospholipids; D018197:Hepatoblastoma / Q000628:therapy*; D006801:Humans; D007275:Injections, Intravenous; D008081:Liposomes; D008099:Liver / Q000201:enzymology*; D008114:Liver Neoplasms, Experimental / Q000628:therapy*; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008855:Microscopy, Electron, Scanning; D010714:Phosphatidylethanolamines; D012855:Sitosterols; D014018:Tissue Distribution; D014162:Transfection / Q000379:methods
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D020404:Glycerophospholipids; D008081:Liposomes; D010714:Phosphatidylethanolamines; D012855:Sitosterols; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; D002784:Cholesterol; D008156:Luciferases; C011015:lyoniside