Construction and testing of gene therapy retroviral vector expressing bacterial cytosine deaminase gene.
A retroviral vector containing gene for bacterial enzyme cytosine deaminase (CD) under the control of viral LTR sequences was constructed and transfected into packaging cell line GP+envAm12. High virus titer producing single cell clone (1 x 10(7) cfu/ml, determined on NIH 3T3 cells) was isolated and used to transfer CD gene into human mammary carcinoma cell lines in vitro. Transduced cells exhibited high sensitivity to the antifungal drug 5-fluorocytosine (5-FC), whereas parental cells did not. Cocultivation of CD-positive and CD-negative parental cells showed bystander effect, dependent on the ratio of CD-positive cells. No enhancement of 5-FC cytotoxicity by leucovorin was observed in cells expressing cytosine deaminase.
Hlavatý
J
J
Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Hlubinová
K
K
Altaner
C
C
D000818:Animals; D002460:Cell Line; D043525:Cytosine Deaminase; D005437:Flucytosine / Q000494:pharmacology; D005472:Fluorouracil / Q000494:pharmacology; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D002955:Leucovorin / Q000494:pharmacology; D051379:Mice; D009700:Nucleoside Deaminases / Q000235:genetics* / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*
D005437:Flucytosine; D009700:Nucleoside Deaminases; D043525:Cytosine Deaminase; D002955:Leucovorin; D005472:Fluorouracil