Asialoganglioside enhances the efficiency of gene transfection mediated by cationic liposomes with a cationic cholesterol derivative.
We investigated the transfection efficiency mediated by asialoganglioside-containing cationic liposomes. Previously we reported that monosialoganglioside GM(1) (GM(1a)) enhanced transfection efficiency. In this study, we investigated the effects of sialic acid in gangliosides on transfection efficiency. Two mammalian culture cell lines HeLa and HepG2 were transfected with luciferase plasmids (pGL3) using cationic liposomes which contain monosialoganglioside GM(1) (GM(1a)) or its asialic counterpart, asialoganglioside GM(1) (GA(1)). Both GM(1a) and GA(1) enhanced the efficiency of transfection mediated by cationic liposomes, and GA(1) exhibited higher efficiency than GM(1a) in both cell lines. Transfection efficiency of ganglioside-containing liposomes was also assessed by the effects of antisense oligonucleotides (AS-ODN) for bcl-2 gene, which suppresses apoptotic cell death. Western blotting analysis revealed that the expression of Bcl-2 was decreased by AS-ODN, and the reduction of protein expression in cells treated with GA(1)-containing liposomes was more remarkable than that with GM(1a)-containing liposomes. Furthermore, the induction rate of apoptosis was higher in cells treated with AS-ODN with GA(1)-containing liposomes. Together with the results obtained by luciferase assay mentioned above, the removal of sialic acid from ganglioside causes the enhancement of efficiency of transfection mediated by cationic liposomes.
Noguchi
Shoichiro
S
Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan.
Hirashima
Naohide
N
Nakanishi
Mamoru
M
D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002412:Cations; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000737:chemistry*; D004337:Drug Carriers; D005690:Galactose / Q000737:chemistry; D018014:Gene Transfer Techniques; D019254:Genes, bcl-2 / Q000235:genetics; D006028:Glycosphingolipids / Q000737:chemistry / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage; D014162:Transfection
D002412:Cations; D004337:Drug Carriers; D006028:Glycosphingolipids; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D002784:Cholesterol; D008156:Luciferases; D005690:Galactose