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Tumor-specific gene expression using regulatory elements of the glucose transporter isoform 1 gene

Cancer Gene Therapy volume 11, pages 41–51 (2004)Cite this article

Abstract

In order to achieve tumor-specific targeting of adeno-associated virus (AAV)-mediated gene expression, the promoter of the glucose transporter isoform 1 (GLUT1) gene was cloned upstream of the enhanced green fluorescence protein (EGFP) and the herpes simplex virus thymidine kinase (HSVtk) gene. FACS analysis performed at 48 h after transient infection with rAAV/cytomegalovirus (CMV)egfp viral particles revealed an increase of fluorescence in all the cell lines tested. However, EGFP expression under control of the GLUT1 promoter element (rAAV/GTI-1.3egfp) was limited to the tumor cells and oncogene-transformed cells. Evidence for phosphorylation of the HSVtk substrates ganciclovir (GCV) and 125I-deoxycytidine was found in all transfected tumor cell lines compared to noninfected controls (HCT116: 111%; MH3924A: 130%; HaCaT-RT3: 257% increase), but not in HaCaT and HUVEC cells. Furthermore, tumor cells and the oncogene-transformed (ras) cell line HaCaT-RT3 showed a GCV-induced reduction in cell number (HCT116: −71%; MH3924A: −43% and HaCaT-RT3: −31%). No statistically relevant cytotoxic effect was observed in HaCaT (6% decrease) and HUVEC cells (2% decrease). Furthermore, a reduction of 3H-thymidine incorporation into the DNA was seen after treatment with GCV (HCT116: 38%; MH3924A: 33% and HaCaT-RT3: 37% decrease). In a therapy study of HSVtk-expressing tumors with GCV, we achieved total tumor remission.

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Acknowledgements

We thank M Mahmut, I Morr and I Preugschat-Gumprecht for their technical help. We also thank U Schierbaum and K Leotta for their help in animal experiments. Supported by the Forschungsförderung of the University of Heidelberg and the Tumorzentrum Heidelberg-Mannheim.

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Authors and Affiliations

  1. Clinical Cooperation Unit Nuclear Medicine, Heidelberg, Germany

    Stephanie Sieger, Shiming Jiang, Helmut Eskerski, Frank Schönsiegel, Annette Altmann & Uwe Haberkorn

  2. Applied Tumor Virology, German Cancer Research Center, Heidelberg, Germany

    Jürgen Kleinschmidt

  3. Department of Nuclear Medicine, University of Heidelberg, Germany

    Walter Mier & Uwe Haberkorn

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  1. Stephanie Sieger
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Correspondence to Uwe Haberkorn.

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Sieger, S., Jiang, S., Kleinschmidt, J. et al. Tumor-specific gene expression using regulatory elements of the glucose transporter isoform 1 gene. Cancer Gene Ther 11, 41–51 (2004). https://doi.org/10.1038/sj.cgt.7700654

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