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In vitro-induced resistance to the deoxycytidine analogues cytarabine (AraC) and 5-aza-2′-deoxycytidine (DAC) in a rat model for acute myeloid leukemia is mediated by mutations in the deoxycytidine kinase (dck) gene

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Abstract

The deoxycytidine kinase (dck) gene encodes the enzyme responsible for the metabolic activation of the antileukemic drugs cytosine arabinoside (AraC) and 5-aza-2′-deoxycytidine (decitabine, DAC). Thedck locus was analyzed at the chromosomal and the molecular level in a model of rat leukemic cell lines, in which AraC and DAC resistance was induced, that was marked bydck deficiency. At the chromosomal level, karyotype analysis of metaphase spreads revealed the presence of an aberrant 2q+ chromosome in the AraC-resistant cell line and a (Xq;11q) translocation in its subclone RA/7. The DAC-resistant lines were identical to the parental RCL/O. Fluorescence in situ hybridization on normal rat fibroblast metaphase spreads localized the ratdck gene to chromosome 14q21-q22, a region that was not involved in any of the observed karyotypic aberrations. Analysis at the molecular level revealed an identical rearrangement of thedck gene in the AraC-resistant cell line RCL/A and its subclone RA/7 that resulted in the absence ofdck expression, as assessed by RT-PCR. No genomic rearrangements were observed in a DAC-resistant cell line RCL/D or in its subclone RD/1. However, detection of a single-stranded conformation polymorphism (SSCP) allowed the identification of a single C to G substitution (His to Gln) in thedck cDNA of the DAC-resistant RD/1 clone. The data demonstrate that exposure to AraC and DAC induces a resistant phenotype marked by functionaldck deficiency that may be the consequence of mutations occurring in thedck gene.

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Authors and Affiliations

  1. Department of Pharmacology, University of North Carolina at Chapel Hill, 1106 FLOB, CB 7365, 27599, Chapel Hill, NC, USA

    A. P. A. Stegmann

  2. Laboratory for Experimental Hematology, Department of Hematology, University Hospital Leiden, The Netherlands

    M. W. Honders, R. Willemze & J. E. Landegent

  3. Department of Cell Biology and Cytogenetics, Erasmus University, Rotterdam, The Netherlands

    A. Hagemeijer

  4. Laboratory of Carcinogenesis and Mutagenesis, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands

    B. Hoebee

Authors
  1. A. P. A. Stegmann
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  2. M. W. Honders
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  3. A. Hagemeijer
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  4. B. Hoebee
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  5. R. Willemze
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  6. J. E. Landegent
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This work was supported by grant KWF 90-02 from the Dutch Cancer Foundation

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Stegmann, A.P.A., Honders, M.W., Hagemeijer, A. et al. In vitro-induced resistance to the deoxycytidine analogues cytarabine (AraC) and 5-aza-2′-deoxycytidine (DAC) in a rat model for acute myeloid leukemia is mediated by mutations in the deoxycytidine kinase (dck) gene. Ann Hematol 71, 41–47 (1995). https://doi.org/10.1007/BF01696231

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  • DOI: https://doi.org/10.1007/BF01696231

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