Drug resistance to 5-aza-2′-deoxycytidine, 2′,2′-difluorodeoxycytidine, and cytosine arabinoside conferred by retroviral-mediated transfer of human cytidine deaminase cDNA into murine cells

被引:81
作者
Eliopoulos, N
Cournoyer, D
Momparler, RL
机构
[1] Hop St Justine, Ctr Rech Pediat, Montreal, PQ H3T 1C5, Canada
[2] Univ Montreal, Dept Pharmacol, Montreal, PQ H3T 1C5, Canada
[3] Montreal Gen Hosp, Dept Med, Montreal, PQ H3G 1A4, Canada
[4] Montreal Gen Hosp, Dept Oncol, Montreal, PQ H3G 1A4, Canada
基金
英国医学研究理事会;
关键词
cytidine deaminase; 5-AZA-2 '-deoxycytidine; 2 ' 2 '-difluorodeoxycytidine; cytosine arabinoside; 3,4,5,6-tetrahydrouridine;
D O I
10.1007/s002800050832
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Purpose: The hematopoietic toxicity produced by the cytosine nucleoside analogs is a critical problem that limits their effectiveness in cancer therapy. One strategy to prevent this dose-limiting toxicity would be to insert a gene for drug resistance to these analogs into normal bone marrow cells. Cytidine (CR) deaminase can deaminate and thus inactivate 5-aza-2'-deoxycytidine (5-AZA-CdR), 2',2'-difluorodeoxycytidine (dFdC) and cytosine arabinoside (ARA-C). The aim of this study was to determine if gene transfer of CR deaminase into murine fibroblast cells confers drug resistance to these cytosine nucleoside analogs and if this resistance can be prevented by the CR deaminase inhibitor, 3,4,5,6-tetrahydrouridine (THU). Methods: NIH 3T3 murine fibroblast cells were transduced with retroviral particles containing the human CR deaminase cDNA. Assays measuring CR deaminase activity as well as the inhibitory action of 5-AZA-CdR, dFdC and ARA-C on colony formation, were performed in the presence of different concentrations of THU. Results: Retroviral-mediated transfer of the CR deaminase gene into 3T3 fibroblasts produced a considerable increase in CR deaminase activity. The transduced cells also showed significant drug resistance to 5-AZA-CdR, dFdC and ARA-C, as demonstrated by a clonogenic assay. This drug resistance phenotype and elevated CR deaminase activity were reversed by THU. Conclusions: These findings indicate that the CR deaminase gene can potentially be used in cancer gene therapy for protecting normal cells against the cytotoxic actions of different cytosine nucleoside analogs. In addition, the CR deaminase-transduced cells can be used as a model for screening different CR deaminase inhibitors in an intact cellular system.
引用
收藏
页码:373 / 378
页数:6
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