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Apoptosis Induction With 5-Fluorocytosine/Cytosine Deaminase gene Therapy for Human Malignant Glioma Cells Mediated by Adenovirus

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Abstract

Previously, we evaluated the therapeutic efficacy of the adenovirus-mediated transduction of the cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) for malignant gliomas. However, the molecular pathways that mediate the 5-FC/CD gene therapy-induced cell death remains to be elucidated. In this study, we examined the induction of apoptosis and the role of caspases in 5-FC/CD gene therapy using human malignant glioma cells [Gli36Δ5 (mutated p53) and U87MG (wild p53)]. The treatment with 5-FC/CD gene-therapy-induced apoptosis both in Gli36Δ5 cells and in U87MG cells according to flow cytometric analysis. Immunoblot analysis revealed that caspases 3 and 9 were processed in response to 5-FC/CD in a concentration- and time-dependent manner, but caspase 8 was not. Each caspase 3 and 9 inhibitor significantly reduced apoptosis triggered by 5-FC/CD, but the caspase 8 inhibitor did not affect apoptosis induction. 5-FC/CD significantly promoted the release of cytochorme c from mitochondria in a concentration-dependent manner. These results indicate that 5-FC/CD gene therapy induces apoptosis in human malignant glioma cells and that the apoptotic cell death is mediated by the activation of mitochondrial caspase cascades involving caspases 3 and 9. This is the first report concerning the apoptotic mechanism of 5-FC/CD gene therapy, and these findings could be used to increase the efficacy of suicide gene therapy systems for the treatment of malignant glioma.

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Authors and Affiliations

  1. Department of Neurological Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama

    Kazuhiko Kurozumi, Takashi Tamiya, Yasuhiro Ono, Shinji Otsuka, Hirokazu Kambara, Yoshiaki Adachi, Tomotsugu Ichikawa & Takashi Ohmoto

  2. Department of Molecular Medicine, Sapporo Medical University, Sapporo, Japan

    Hirofumi Hamada

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  1. Kazuhiko Kurozumi
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  2. Takashi Tamiya
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  3. Yasuhiro Ono
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  4. Shinji Otsuka
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  5. Hirokazu Kambara
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Kurozumi, K., Tamiya, T., Ono, Y. et al. Apoptosis Induction With 5-Fluorocytosine/Cytosine Deaminase gene Therapy for Human Malignant Glioma Cells Mediated by Adenovirus. J Neurooncol 66, 117–127 (2004). https://doi.org/10.1023/B:NEON.0000013494.98345.80

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