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Down-regulation of Wilms’ tumor 1 expression in glioblastoma cells increases radiosensitivity independently of p53

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Abstract

The Wilms’ tumor 1 (WT1) gene is overexpressed in human glioblastoma and correlates with wild-type p53 status. In other cell types, WT1 inhibits p53-mediated apoptosis in response to DNA damaging agents. However, neither this interaction nor the relationship between WT1 and radiosensitivity has been studied in glioblastoma. To study this interaction, we generated LN-229 glioma cell lines (p53 mutant) stably expressing WT1 isoforms and induced apoptosis by transfecting with different doses of wild-type p53 plasmid expression vector. Constitutive expression of WT1 did not protect against exogenous p53-mediated apoptosis. Likewise, WT1 expression did not protect against endogenous p53-mediated cell death induced by radiotherapy in U87MG cells, which contain functional wild-type p53. We then tested the efficacy of WT1 siRNA in inhibiting WT1 expression and its effect on radiosensitivity. In T98G and LN-18 glioma cells, which possess p53 mutations, WT1 siRNA decreased WT1 protein to almost undetectable levels by 96-h post-transfection. Furthermore, WT1 siRNA transfection caused a significantly larger decrease in viability following irradiation than was seen in untransfected cells in both cell lines after treatment with ED50 of ionizing radiation. In conclusion, WT1 overexpression did not protect against p53-mediated apoptosis or ionizing radiation induced cell death. WT1 siRNA increased the radiosensitivity of two human glioma cell lines independently of p53. Anti-WT1 strategies may, therefore, prove useful in improving the response of glioblastoma to radiotherapy, thus potentially improving patient survival.

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Abbreviations

WT1:

Wilms’ tumor 1

siRNA:

Short interfering RNA

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Acknowledgments

We thank Dr. Jessica McCready for critically reading the manuscript. We also thank Dr. Charles T. Roberts, Jr. and Dr. Sumitra Deb for generously providing WT1 and p53 plasmids. The research presented in this paper was supported in part by a Medical Student Summer Fellowship from the American Brain Tumor Association and by the Hord and Cullather funds of the MCV Foundation.

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Authors and Affiliations

  1. Department of Neurosurgery, Virginia Commonwealth University, Medical College of Virginia Campus, P.O. Box 980631, Richmond, VA, 23298-0631, USA

    Aaron J. Clark, Mike Y. Chen, Helen Fillmore, Wagner G. Dos Santos, Timothy E. Van Meter, Martin R. Graf & William C. Broaddus

  2. Department of Anatomy and Neurobiology, Virginia Commonwealth University, Medical College of Virginia Campus, Richmond, VA, USA

    Aaron J. Clark, Dana C. Chan, Mike Y. Chen, Helen Fillmore & William C. Broaddus

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  1. Aaron J. Clark
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  8. William C. Broaddus
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Correspondence to William C. Broaddus.

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Clark, A.J., Chan, D.C., Chen, M.Y. et al. Down-regulation of Wilms’ tumor 1 expression in glioblastoma cells increases radiosensitivity independently of p53. J Neurooncol 83, 163–172 (2007). https://doi.org/10.1007/s11060-006-9317-8

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  • DOI: https://doi.org/10.1007/s11060-006-9317-8

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