Functional characterization of an EGF receptor with a truncated extracellular domain expressed in glioblastomas with EGFR gene amplification

A. Jonas Ekstrand, Nicola Longo, Melissa L. Hamid, Jeffrey J. Olson, Lu Liu, V. Peter Collins, C. David James*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

252 Scopus citations

Abstract

The most common type of alteration of the epidermal growth factor receptor gene (EGFR) in human glioblastomas results in the synthesis of an aberrant mRNA lacking 801 bases that encode amino acids 6-273 of the receptor's extracellular domain. To study the effects of this mutation on receptor function, we have developed chinese hamster ovary cell transfectants which express the mutant EGF receptor. Comparison of wild-type and mutant receptor properties in this cell host indicates that the truncated receptor does not bind EGF or TGF-α and, consequently, DNA synthesis is not stimulated in cultures of mutant transfectants by either grown factor. However, levels of DNA synthesis determined for mutant transfectants in serum-free media are several-fold higher than those determined for corresponding cultures of wild-type transfectants. Western blot analysis with anti-phosphotyrosine antibody indicates that the mutant receptor is constitutively phosphorylated in CHO cells, and the same analysis applied to lysates of glioblastoma biopsies reveals the altered receptor is readily detectable as a phosphotyrosine protein in tumors for which there is evidence of corresponding EGFR gene and transcript alterations. In total, these results indicate that the aberrant EGF receptor synthesized in glioblastomas, and which lacks a portion of the extracellular domain necessary for ligand binding, is an activated tyrosine kinase.

Original languageEnglish (US)
Pages (from-to)2313-2320
Number of pages8
JournalOncogene
Volume9
Issue number8
StatePublished - Aug 1994

Funding

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

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