Liposomal delivery of the herpes simplex virus thymidine kinase gene in glioma: improvement of cell sensitization to ganciclovir.
In this study, we investigated whether the regulation and the copy number of the herpes simplex virus thymidine kinase (HSVtk) gene increased the sensitization to ganciclovir (GCV) of glioma cell lines (Rat C6 and human U118-MG) using liposome-mediated gene transfer. Three recombinant plasmids carrying the HSVtk gene driven by the thymidine kinase promoter in single (pAGo) and double copy (pYED) or by the human cytomegalovirus promoter (pCMVtk) were used for the transfection. The DNA delivery was optimized by screening a panel of cationic liposomes using Lac-Z and luciferase as reporter genes. The efficiency of transfection reached 33% to 36% in vitro but only 18.6% in vivo after an intratumoral injection of DNA-liposome complexes. Moreover, after transfection of the three plasmids, the cell-killing effect of GCV was evaluated. A significant enhancement (four- to fivefold) of the cell sensitivity to GCV was shown in pCMVtk and pYED as compared with pAGo-transfected cells in both cell lines. According to the plasmid, the effect of the HSVtk/GCV system was confirmed by in vivo experiments and was objectified by a higher tumor weight reduction with pCMVtk (49%) than pAGo (27%). From these results, we conclude that (1) the gene transfer can be achieved by cationic liposomes both in vitro and in vivo and that (2) using this type of vector, the antitumor effect of the HSVtk/GCV system could be potentiated by the up-regulation of HSVtk gene duplication.
Zerrouqi
A
A
Laboratory of Medical Oncology Department, Pitié-Salpêtrière Hospital, Paris, France.
Rixe
O
O
Ghoumari
A M
AM
Yarovoi
S V
SV
Mouawad
R
R
Khayat
D
D
Soubrane
C
C
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D005434:Flow Cytometry; D015774:Ganciclovir / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000378:metabolism*; D009374:Neoplasms, Experimental / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D051381:Rats; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000235:genetics; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D008081:Liposomes; D010714:Phosphatidylethanolamines; C020888:1,2-dielaidoylphosphatidylethanolamine; D013937:Thymidine Kinase; D001616:beta-Galactosidase; D015774:Ganciclovir