Gene silencing for epidermal growth factor receptor variant III induces cell-specific cytotoxicity

Farnaz Yamoutpour, Vidya Bodempudi, Shay E. Park, Weihong Pan, Mary Jean Mauzy, Robert A Kratzke, Arkadiusz Z Dudek, David A Potter, Richard A. Woo, Donald M. O'Rourke, Donald J. Tindall, Faris Farassati

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27 Scopus citations

Abstract

Epidermal growth factor receptor variant III (EGFRvIII) is a constitutively active mutant form of EGFR that is expressed in 40% to 50% of gliomas and several other malignancies. Here, we describe the therapeutic effects of silencing EGFRvIII on glioma cell lines in vitro and in vivo. A small interfering RNA molecule against EGFRvIII was introduced into EGFRvIII-expressing glioma cells (U87Δ) by electroporation resulting in complete inhibition of expression of EGFRvIII as early as 48 h post-treatment. During EGFRvIII silencing, a decrease in the proliferation and invasiveness of U87Δ cells was accompanied by an increase in apoptosis (P < 0.05). Notably, EGFRvIII silencing inhibited the signal transduction machinery downstream of EGFRvIII as evidenced by decreases in the activated levels of Ras and extracellular signal-regulated kinase. A lentivirus capable of expressing anti-EGFRvIII short hairpin RNA was also able to achieve progressive silencing of EGFRvIII in U87Δ cells in addition to inhibiting cell proliferation, invasiveness, and colony formation in a significant manner (P < 0.05). Silencing EGFRvIII in U87Δ cultures with this virus reduced the expression of factors involved in epithelial-mesenchymal transition including N-cadherin, β-catenin, Snail, Slug, and paxillin but not E-cadherin. The anti-EGFRvIII lentivirus also affected the cell cycle progression of U87Δ cells with a decrease in G1 and increase in S and G2 fractions. In an in vivo model, tumor growth was completely inhibited in severe combined immunodeficient mice (n = 10) injected s.c. with U87Δ cells treated with the anti-EGFRvIII lentivirus (P = 0.005). We conclude that gene specific silencing of EGFRvIII is a promising strategy for treating cancers that contain this mutated receptor.

Original languageEnglish (US)
Pages (from-to)3586-3597
Number of pages12
JournalMolecular Cancer Therapeutics
Volume7
Issue number11
DOIs
StatePublished - Nov 1 2008

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    Yamoutpour, F., Bodempudi, V., Park, S. E., Pan, W., Mauzy, M. J., Kratzke, R. A., Dudek, A. Z., Potter, D. A., Woo, R. A., O'Rourke, D. M., Tindall, D. J., & Farassati, F. (2008). Gene silencing for epidermal growth factor receptor variant III induces cell-specific cytotoxicity. Molecular Cancer Therapeutics, 7(11), 3586-3597. https://doi.org/10.1158/1535-7163.MCT-08-0653