Escherichia coli gpt gene sensitizes rat glioma cells to killing by 6-thioxanthine or 6-thioguanine.
Genes that encode enzymes that convert inactive "prodrugs" into anticancer metabolites may be therapeutically useful against brain tumors. Unlike other genes tested to date in brain tumor models, the Escherichia coli gpt gene is unique in that it not only sensitizes cells to the prodrug 6-thioxanthine (6TX) but also encodes resistance to a different regimen (mycophenolic acid, xanthine, and hypoxanthine), thus providing a means to select for gpt-positive cells. In the present study, rat C6 glioma cells were infected with a retrovirus vector that transduces this gene. A clonal line (C6GPT-7) was derived that exhibited significant 6TX susceptibility in vitro with an ID50 of 2.5 mumol/L, whereas 50% growth inhibition of parental C6 cells was not achieved at concentrations tested (up to 50 mumol/L). This line also exhibited significant sensitivity to 6-thioguanine (6TG), with an ID50 of 0.05 mumol/L, whereas 50% growth inhibition of parental C6 cells was achieved at 0.5 mumol/L. In a "bystander" assay, C6GPT-7 tumor cells efficiently transferred 6TX sensitivity to C6 cells at ratios as low as 1:9 (C6GPT-7:C6). This in vitro bystander effect was abrogated when C6GPT-7 and C6 cells were separated by a microporous membrane, suggesting that it was not mediated by highly diffusible metabolites. In vivo both 6TX and 6TG significantly inhibited the growth of subcutaneously transplanted C6GPT-7 cells but not that of C6 cells in athymic mice. In an intracerebral model, both 6TX and 6TG exhibited significant antiproliferative effects against tumors formed by C6GPT-7 cells. These findings provide a basis for exploring further gene therapy strategies based on in vivo transfer of the E coli gpt gene to provide chemosensitivity against 6TX and 6TG.
Tamiya
T
T
Department of Surgery, Massachusetts General Hospital, Boston 02129, USA.
Ono
Y
Y
Wei
M X
MX
Mroz
P J
PJ
Moolten
F L
FL
Chiocca
E A
EA
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000633:toxicity*; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005910:Glioma / Q000188:drug therapy / Q000473:pathology*; D007041:Hypoxanthine Phosphoribosyltransferase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013866:Thioguanine / Q000633:toxicity*; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014183:Transplantation, Heterologous; D014970:Xanthines / Q000633:toxicity*
D000964:Antimetabolites, Antineoplastic; D011994:Recombinant Proteins; D014970:Xanthines; C054700:6-thioxanthine; D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine