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  • Experimental Therapeutics
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Induction of DNA breaks and apoptosis in crosslink-hypersensitive V79 cells by the cytostatic drug β-D-glucosyl-ifosfamide mustard

British Journal of Cancer volume 86, pages 130–135 (2002)Cite this article

Abstract

To study molecular aspects of cytotoxicity of the anticancer drug β-D-glucose-ifosfamide mustard we investigated the potential of the agent to induce apoptosis and DNA breakage. Since β-D-glucose-ifosfamide mustard generates DNA interstrand crosslinks, we used as an in vitro model system a pair of isogenic Chinese hamster V79 cells differing in their sensitivity to crosslinking agents. CL-V5B cells are dramatically more sensitive (30-fold based on D10 values) to the cytotoxic effects of β-D-glucose-ifosfamide mustard as compared to parental V79B cells. After 48 h of pulse-treatment with the agent, sensitive cells but not the resistant parental line undergo apoptosis and necrosis, with apoptosis being the predominant form of cell death (70 and 20% of apoptosis and necrosis, respectively). Apoptosis increased as a function of dose and was accompanied by induction of DNA double-strand breaks in the hypersensitive cells. Furthermore, a strong decline in the level of Bcl-2 protein and activation of caspases-3, -8 and -9 were observed. The resistant parental cells were refractory to all these parameters. Bcl-2 decline in the sensitive cells preceded apoptosis, and transfection-mediated overexpression of Bcl-2 protected at least in part from apoptosis. From the data we hypothesize that non-repaired crosslinks induced by β-D-glucose-ifosfamide mustard are transformed into double-strand breaks which trigger apoptosis via a Bcl-2 dependent pathway.

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Acknowledgements

This work was supported by Deutsche Forschungsgemeinschaft, DFG Ka756/4-4, Rheinland-Pfalt SFB 519/B4, and Stiftung.

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Authors and Affiliations

  1. Division of Applied Toxicology, Institute of Toxicology, University of Mainz, Obere Zahlbacher Str, Mainz, 67, D-55131, Germany

    R Becker, A Ritter, U Eichhorn, J Lips & B Kaina

  2. Division of Molecular Toxicology, German Cancer Research Center, Im Neuenheimer Feld 280, Heidelberg, D-69120, Germany

    B Bertram & M Wiessler

  3. Department of Radiation Genetics and Chemical Mutagenesis, Leiden University Medical Center, Wassenaarseweg, Leiden, 72, NL-2333, The Netherlands

    M Z Zdzienicka

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  1. R Becker
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  2. A Ritter
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  3. U Eichhorn
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  4. J Lips
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Correspondence to B Kaina.

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Becker, R., Ritter, A., Eichhorn, U. et al. Induction of DNA breaks and apoptosis in crosslink-hypersensitive V79 cells by the cytostatic drug β-D-glucosyl-ifosfamide mustard. Br J Cancer 86, 130–135 (2002). https://doi.org/10.1038/sj.bjc.6600027

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  • DOI: https://doi.org/10.1038/sj.bjc.6600027

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