Chordoma, the molecular hallmark of which is T (
brachyury), is a rare malignant bone tumour with a high risk of local recurrence and a tumour from which metastatic disease is a common late event. Currently, there is no effective
drug therapy for treating
chordomas, although there is evidence that some patients respond to the empirical use of
epidermal growth factor receptor (EGFR) antagonists. The aim of this study was to determine the role of EGFR in the pathogenesis of
chordoma.
Paraffin-embedded material from 173
chordomas from 160 patients [sacro-coccygeal (n = 94), skull-based (n = 50), and mobile spine (n = 16)] was analysed by immunohistochemistry and revealed total EGFR expression in 69% of cases analysed. Of 147 informative
chordomas analysed by FISH, 38% revealed high-level EGFR polysomy, 4% high-level polysomy with focal amplification, 18% low-level polysomy, and 39% disomy. Phospho-
receptor tyrosine kinase array membranes showed EGFR activation in the
chordoma cell line U-CH1 and all of the three
chordomas analysed. Direct sequencing of EGFR (exons 18-21), KRAS, NRAS, HRAS (exons 2, 3), and BRAF (exons 11, 15) using
DNA from 62
chordomas failed to reveal mutations. PTEN expression was absent by immunohistochemistry in 19 of 147 (13%) analysed
chordomas, only one of which revealed high-level polysomy of EGFR. The EGFR inhibitor
tyrphostin (AG 1478) markedly inhibited proliferation of the
chordoma cell line U-CH1 in vitro and diminished EGFR phosphorylation in a dose-dependant manner, a finding supported by inhibition of phosphorylated Erk1/2. p-Akt was suppressed to a much lesser degree in these experiments. There was no reduction of T as assessed by western blotting. These data implicate aberrant EGFR signalling in the pathogenesis of
chordoma. This study provides a strategy for patient stratification for treatment with EGFR antagonists.