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Mechanisms of thymidine kinase/ganciclovir and cytosine deaminase/ 5-fluorocytosine suicide gene therapy-induced cell death in glioma cells

Oncogene volume 24, pages 1231–1243 (2005)Cite this article

Abstract

Suicide gene transfer using thymidine kinase (TK) and ganciclovir (GCV) treatment or the cytosine deaminase (CD)/5-fluorocytosine (5-FC) system represents the most widely used approach for gene therapy of cancer. However, molecular pathways and resistance mechanisms remain controversial for GCV-mediated cytotoxicity, and are virtually unknown for the CD/5-FC system. Here, we elucidated some of the cellular pathways in glioma cell lines that were transduced to express the TK or CD gene. In wild-type p53-expressing U87 cells, exposure to GCV and 5-FC resulted in a weak p53 response, although apoptosis was efficiently induced. Cell death triggered by GCV and 5-FC was independent of death receptors, but accompanied by mitochondrial alterations. Whereas expression of Bax remained unaffected, in particular, GCV and also 5-FC caused a decline in the level of Bcl-2. Similar findings were obtained in 9L and T98G glioma cells that express mutant p53, and also underwent mitochondrial apoptosis in both the TK/GCV and CD/5-FC system. Upon treatment of 9L cells with 5-FC, Bcl-xL expression slowly declined, whereas exposure to GCV resulted in the rapid proapoptotic phosphorylation of Bcl-xL. These data suggest that TK/GCV- and CD/5-FC-induced apoptosis does neither require p53 nor death receptors, but converges at a mitochondrial pathway triggered by different mechanisms of modulation of Bcl-2 proteins.

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Acknowledgements

This study was supported by grants from the BioRegio program of the German Ministry of Research and Technology (Forschungsverbund Somatische Gentherapie), the Deutsche Forschungsgemeinschaft (DFG Kr1711/1-1), and the Elterninitiative Kinderkrebsklinik Düsseldorf e.V.

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Authors and Affiliations

  1. Institute of Molecular Medicine, Heinrich-Heine-University, Düsseldorf, Germany

    Ute Fischer & Klaus Schulze-Osthoff

  2. Department of Pediatric Hematology, Oncology, and Immunology, Heinrich-Heine-University, Düsseldorf, Germany

    Ute Fischer, Sabine Steffens, Susanne Frank & Christof M Kramm

  3. Department of Neurological Science, University of Liverpool, Liverpool, UK

    Nikolai G Rainov

  4. The Walton Centre for Neurology and Neurosurgery, NHS Trust, Liverpool, UK

    Nikolai G Rainov

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  1. Ute Fischer
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  2. Sabine Steffens
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  4. Nikolai G Rainov
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  5. Klaus Schulze-Osthoff
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Correspondence to Klaus Schulze-Osthoff.

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Fischer, U., Steffens, S., Frank, S. et al. Mechanisms of thymidine kinase/ganciclovir and cytosine deaminase/ 5-fluorocytosine suicide gene therapy-induced cell death in glioma cells. Oncogene 24, 1231–1243 (2005). https://doi.org/10.1038/sj.onc.1208290

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