Kawaura C, Hasegawa S, Hirashima N, Nakanishi M
Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
Biol Pharm Bull. 2000 Jun;23(6):778-80. doi: 10.1248/bpb.23.778.
We have studied the effects of monosialoganglioside (GM1)-containing cationic liposomes with a cationic cholesterol on the liposome-mediated gene transfection into mammalian culture cells. The results showed that both cationic liposomes with either a cationic cholesterol derivative of a hydrophobic amino head group (I) and a hydrophilic amino head group (II) promoted the transfection of luciferase plasmids (pGL3) into HeLa and CHO-K1 cells more than the control cationic liposomes without GM1. In addition, we found that cationic liposomes with a cationic cholesterol derivative (II) were about ten times as effective as that by commercially available cationic liposome Lipofectin. Confocal fluorescence microscopy showed that the liposome/DNA complex was transferred more efficiently into the target cells by the GM1-containing liposomes than by the liposomes without GM1. In proportion to the above results, free antisense DNAs were also more efficiently transferred into the nucleus of the target cells by the GM1-containing liposomes. When there was 100 mM galactose in the transfection medium, the luciferase activity by the GM1-containing liposomes was reduced to the level of the control liposomes. The results suggest that GM1-containing cationic liposomes with a cationic cholesterol derivative of a hydrophobic amino head group or a hydrophilic amino head group should significantly increase the transfection efficiency of plasmid DNAs and antisense DNAs by galactose receptor-mediated endocytosis. This means that the GM1-containing liposomes described here should be very promising for gene transfection in vitro.
我们研究了含有单唾液酸神经节苷脂（GM1）的阳离子脂质体与阳离子胆固醇对脂质体介导的基因转染至哺乳动物培养细胞的影响。结果表明，带有疏水性氨基头基（I）和亲水性氨基头基（II）的阳离子胆固醇衍生物的两种阳离子脂质体，比不含GM1的对照阳离子脂质体更能促进荧光素酶质粒（pGL3）转染至HeLa和CHO-K1细胞。此外，我们发现带有阳离子胆固醇衍生物（II）的阳离子脂质体的效果约为市售阳离子脂质体Lipofectin的十倍。共聚焦荧光显微镜显示，含GM1的脂质体比不含GM1的脂质体更有效地将脂质体/DNA复合物转运至靶细胞。与上述结果一致，含GM1的脂质体也能更有效地将游离反义DNA转运至靶细胞的细胞核。当转染培养基中有100 mM半乳糖时，含GM1的脂质体的荧光素酶活性降至对照脂质体的水平。结果表明，带有疏水性氨基头基或亲水性氨基头基的阳离子胆固醇衍生物的含GM1阳离子脂质体应能通过半乳糖受体介导的内吞作用显著提高质粒DNA和反义DNA的转染效率。这意味着此处所述的含GM1脂质体在体外基因转染方面应极具前景。
