Preparation of a TK/GCV administration system mediated by transferrin modified pro-cationic liposomes.
Transferrin modified pro-cationic liposomes were prepared and used to investigate the effect of targeting therapeutic genes to human hepatoma carcinoma cells in vitro. The main lipid CHETA, cholest-5-en-3beta-yl[2-[[4-[(carboxymethyl)dithio]-1-iminobutyl]amino]ethyl] carbamate (C36H61N3O4S2), was synthesized and used to prepare pro-cationic liposomes. The thymidine kinase (TK) gene loaded pro-cationic liposomes were prepared by first mixing the plasmid DNA and protamine together, and then incubating the resulted polyplexes with blank pro-cationic liposomes preformed by the thin film dispersion-sonication method. Transferrin (Tf) was adsorbed on the surface of pro-cationic liposomes via electrostatic interactions to form transferrin modified pro-cationic liposomes. Cellular association was measured by fluorimetry at excitation and emission wavelengths of 490 and 520 nm, respectively. The viability of TK gene infected cells following administration of ganciclovir (GCV) was investigated by MTT assay. The transferrin modified TK gene pro-cationic liposomes had a mean diameter of 240 +/- 12 nm and zeta potential of -24.10 +/- 2.5 mV (n = 3). The transmission electron microscopy image indicated that most of the liposomes were relatively regular and spherical with a condensed core inside. Cell-associated fluorescence of Tf-liposomes and unmodified liposomes (without transferrin) was 7.8 x 10(6), and 3.2 x 10(6) per milligram protein, respectively. Compared to Lipofectamine 2000 (Invitrogen, USA) the pro-cationic liposomes and transferrin modified pro-cationic liposomes had less cytotoxicity to cells. The transduced TK gene HepG2 cells were more sensitive to GCV than the un-transduced TK gene ones and the human normal Chang liver cells were not affected by the TK/GCV system mediated by procationic liposomes.
Zhong
Zhi-Rong
ZR
West China School of Pharmacy, Sichuan University, Chengdu, Sichuan, PR China.
Zhang
Zhi-Rong
ZR
Deng
Yong
Y
Liu
Ji
J
Song
Qing-Guo
QG
Liu
Jie
J
He
Qin
Q
D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002412:Cations / Q000737:chemistry; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage*; D004229:Dithiothreitol / Q000737:chemistry; D004337:Drug Carriers; D004563:Electrochemistry; D015774:Ganciclovir / Q000737:chemistry*; D006801:Humans; D007202:Indicators and Reagents; D008081:Liposomes / Q000737:chemistry*; D013439:Sulfhydryl Reagents / Q000737:chemistry; D013778:Tetrazolium Salts; D013844:Thiazoles; D013937:Thymidine Kinase / Q000235:genetics*; D014168:Transferrin / Q000737:chemistry*
D002412:Cations; D004337:Drug Carriers; D007202:Indicators and Reagents; D008081:Liposomes; D013439:Sulfhydryl Reagents; D013778:Tetrazolium Salts; D013844:Thiazoles; D014168:Transferrin; D004247:DNA; D013937:Thymidine Kinase; C022616:thiazolyl blue; D015774:Ganciclovir; D004229:Dithiothreitol