A human brain tumor-derived PDGFR-alpha deletion mutant is transforming.
Aberrant receptor tyrosine kinase signaling plays an important role in the molecular pathogenesis of brain tumors. We have been studying a previously identified human glioblastoma-derived PDGFR-alpha mutant that has an in-frame deletion in the extracellular domain, causing loss of exons 8 and 9 (PDGFR-alpha(delta8,9)). In the primary tumor, this deletion mutant receptor was shown to be amplified and overexpressed. The purpose of this study was to determine the expression, activity, localization, and transformation properties of this deletion mutant. In the absence of serum, or PDGF-AA, PDGFR-alpha(delta8,9) was phosphorylated on tyrosine residues, indicating ligand-independent autoactivation. Localization by staining and cell surface biotinylation studies revealed expression of the deletion mutant predominantly in the cytoplasm, with very little present on the cell surface. To determine if PDGFR-alpha(delta8,9) was oncogenic, we transfected wild-type and mutant receptors into Rat1 cells and performed analyses of cell growth, in vitro transformation, and subcutaneous growth in the nude mouse. PDGFR-alpha(delta8,9)-expressing cells displayed enhanced cell growth and survival in low serum, and formed foci in monolayer cultures. PDGFR-alpha(delta8,9)-expressing Rat1 cells were also tumorigenic when injected subcutaneously into nude mice. Expression of PDGFR-alpha(delta8,9) was also associated with increased c-Jun phosphorylation in the absence of PDGF ligand, demonstrating also that the mutant receptor is associated with altered intracellular signaling. These data demonstrate that PDGFR-alpha(delta8,9) is transforming, and it is the first demonstration of a naturally occurring tumor-derived mutant PDGFR-alpha with oncogenic properties.
Clarke
I D
ID
Arthur and Sonia Labatt Brain Tumor Research Laboratory, Hospital for Sick Children, University of Toronto, Ontario, Canada.
Dirks
P B
PB
D000818:Animals; D001932:Brain Neoplasms / Q000235:genetics*; D019556:COS Cells; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003593:Cytoplasm / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D010766:Phosphorylation; D016755:Proto-Oncogene Proteins c-jun / Q000502:physiology; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000235:genetics* / Q000502:physiology; D017384:Sequence Deletion
D008024:Ligands; D016755:Proto-Oncogene Proteins c-jun; D020796:Receptor, Platelet-Derived Growth Factor alpha