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RNA interference against a glioma-derived allele of EGFR induces blockade at G2M

Oncogene volume 24, pages 829–837 (2005)Cite this article

Abstract

Amplification and mutation of the epidermal growth factor receptor (EGFR) is common in astrocytoma. The most frequently occurring mutation (ΔEGFR, EGFRvIII) deletes exons 2–7 from this receptor tyrosine kinase (RTK), and signals constitutively in the absence of ligand. ΔEGFR is not found in normal tissue, and therefore represents an attractive therapeutic target. Here, we show that a small interfering RNA (siRNA) directed against the unique exon 1/exon 8 junction sequence of ΔEGFR efficiently suppressed expression of ΔEGFR in rodent fibroblasts and in two human glioblastoma cell lines. SiRNA-mediated depletion of ΔEGFR led to reduction in the levels of phosphorylated Akt in glioma cells, was associated with increased apoptosis, and induced partial arrest at the G2M phase of the cell cycle. Inhibitors of PI3 kinase cooperated with siRNA treatment, leading to further increases in both cell cycle blockade and apoptosis. Importantly, cell cycle blockade could be reversed, and apoptosis rescued using a conditional allele of Akt, implicating Akt as a primary target of combination therapy. This study demonstrates the therapeutic potential of siRNA to impact ΔEGFR as a glioma-specific target, and offers a mechanistic rationale for combining siRNA and small molecule inhibitor therapies against distinct components in the EGFR signaling pathway.

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Acknowledgements

We are grateful to Lou Chesler, Steven P Miller, David Morgan, and Kevan Shokat for useful discussions, to Kimberly Specht, Chao Zhang, Kevan Shokat, and Martin McMahon for reagents, and to Chris Hackett, Martin McMahon, and Russ Pieper for critical review of this manuscript. This work was supported by Grants U01CA84290, 1P50CA097257-01, and by a grant from the Brain Tumor Society.

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Authors and Affiliations

  1. Department of Neurology, University of California, San Francisco, CA, USA

    Qi-Wen Fan & William A Weiss

  2. Department of Pediatrics, University of California, San Francisco, CA, USA

    Qi-Wen Fan & William A Weiss

  3. Department of Neurological Surgery, University of California, San Francisco, CA, USA

    Qi-Wen Fan & William A Weiss

  4. Brain Tumor Research Center, University of California, San Francisco, CA, USA

    Qi-Wen Fan & William A Weiss

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  1. Qi-Wen Fan
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Correspondence to William A Weiss.

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Fan, QW., Weiss, W. RNA interference against a glioma-derived allele of EGFR induces blockade at G2M. Oncogene 24, 829–837 (2005). https://doi.org/10.1038/sj.onc.1208227

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