Open AccessHighly efficient suicide gene expression in hepatocellular carcinoma cells by Epstein-Barr virus-based plasmid vectors combined with polyamidoamine dendrimer
Author(s) -
Yoshinori Harada,
Masaki Iwai,
Saiyu Tanaka,
Takeshi Okanoue,
Kei Kashima,
Hiroko Maruyama-Tabata,
Hideyo Hirai,
Etsuko Satoh,
Jirô Imanishi,
Osam Mazda
Publication year - 2000
Publication title -
cancer gene therapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.535
H-Index - 86
eISSN - 1476-5500
pISSN - 0929-1903
DOI - 10.1038/sj.cgt.7700079
Subject(s) - transfection , suicide gene , thymidine kinase , plasmid , herpes simplex virus , genetic enhancement , microbiology and biotechnology , virology , ganciclovir , viral vector , vector (molecular biology) , gene delivery , virus , biology , hepatitis b virus , oncolytic virus , cationic liposome , cell culture , cancer research , gene , recombinant dna , human cytomegalovirus , genetics
The present study was aimed at devising an efficient nonviral strategy for suicide gene therapy of hepatocellular carcinoma (HCC). To improve the efficiency of DNA delivery and expression, we applied Epstein-Barr virus (EBV)-based plasmid vectors instead of conventional plasmid vectors and combined them with cationic liposome (EBV/lipoplex) or polyamidoamine dendrimer (PAAD) (EBV/polyplex). When the beta-galactosidase gene was transferred to HuH7, PLC/PRF/5, or HLE cells, < or =50-fold higher beta-galactosidase activities were demonstrated in the cells transfected with EBV vector compared with those transfected with conventional plasmid vectors. PAAD-mediated transfection of HCC with pSES.Tk (an EBV-based vector carrying the herpes simplex virus-1 thymidine kinase gene) resulted in a marked reduction in viable cell number by the addition of ganciclovir (GCV). The HCC cells transfected with pSES.Tk/PAAD showed 100- to 1000-fold higher susceptibilities to GCV than those transfected with pS.Tk (a conventional plasmid vector carrying herpes simplex virus-1 thymidine kinase gene)/PAAD. The pSES.Tk-transfected HCC cells were effectively killed by day 9 in culture with a clinically feasible concentration of GCV (25 microM), whereas the pS.Tk-transfected cells survived the culture. These results demonstrate highly efficient suicide gene transfer into various HCC cells by EBV-based plasmid vectors in vitro, suggesting the possible application of this nonviral vector system to gene therapy of HCC.