Enhanced tumorigenic behavior of glioblastoma cells expressing a truncated epidermal growth factor receptor is mediated through the Ras-Shc-Grb2 pathway

J Biol Chem. 1996 Oct 11;271(41):25639-45. doi: 10.1074/jbc.271.41.25639.

Abstract

A mutant epidermal growth factor receptor (DeltaEGFR) containing a deletion of 267 amino acids from the extracellular domain is common in human glioblastomas. We have previously shown that the mutant receptor fails to bind EGF, is constitutively phosphorylated, and confers upon U87MG glioblastoma cells expressing it (U87MG. DeltaEGFR), an increased ability to form tumors in mice. Here we demonstrate that the constitutively phosphorylated DeltaEGFR enhances growth of glioblastoma cells through increased activity of Ras: 1) there was an increase in the proportion of Ras present in the GTP-bound form, and 2) introduction of neutralizing anti-Ras 259 antibodies into U87MG and U87MG.DeltaEGFR cells by microinjection inhibited DNA synthesis to the same low level in both cell populations. We also show that the truncated EGF receptor constitutively associates with the adapter proteins Shc and Grb2 which are involved in the recruitment of Ras to activated receptors. Several derivatives of DeltaEGFR containing single, or multiple mutations at critical autophosphorylation sites were constructed and used to demonstrate that the major Shc binding site is Tyr-1148, and that Grb2 association occurs primarily through Tyr-1068. We conclude that the increased tumorigenic potential of glioblastoma cells expressing the truncated EGF receptor is due at least in part to Ras activation presumably involving the Shc and Grb2 adapter proteins.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Adaptor Proteins, Vesicular Transport*
  • Animals
  • Antibodies, Monoclonal
  • Cell Division
  • Cell Line
  • DNA, Neoplasm / biosynthesis
  • ErbB Receptors / biosynthesis*
  • GRB2 Adaptor Protein
  • Glioblastoma / metabolism
  • Glioblastoma / pathology*
  • Guanosine Diphosphate / metabolism
  • Guanosine Triphosphate / metabolism
  • Humans
  • Immunoglobulin G
  • Mice
  • Mice, Nude
  • Mutagenesis, Site-Directed
  • Phosphoproteins / isolation & purification
  • Phosphoproteins / metabolism
  • Point Mutation
  • Proteins / immunology
  • Proteins / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / biosynthesis
  • Sequence Deletion
  • Shc Signaling Adaptor Proteins
  • Signal Transduction*
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Transfection
  • Transplantation, Heterologous
  • Tumor Cells, Cultured
  • ras Proteins / immunology
  • ras Proteins / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Adaptor Proteins, Vesicular Transport
  • Antibodies, Monoclonal
  • DNA, Neoplasm
  • GRB2 Adaptor Protein
  • GRB2 protein, human
  • Grb2 protein, mouse
  • Immunoglobulin G
  • Phosphoproteins
  • Proteins
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • SHC1 protein, human
  • Shc Signaling Adaptor Proteins
  • Shc1 protein, mouse
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Guanosine Diphosphate
  • Guanosine Triphosphate
  • ErbB Receptors
  • ras Proteins