Construction and testing of gene therapy retroviral vector expressing bacterial cytosine deaminase gene

Hlavatý, J.; Hlubinová, K.; Altaner, C.

Neoplasma 46(5): 267-276


ISSN/ISBN: 0028-2685
PMID: 10665841
Accession: 045631093

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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.