title: Proteomic signatures of epidermal growth factor receptor and survival signal pathways correspond to gefitinib sensitivity in head and neck cancer.
abstract: OBJECTIVE
  Gefitinib targeting of the epidermal growth factor receptor (EGFR) has shown limited activity in clinical trials of head and neck squamous cell carcinoma (HNSCC). To investigate the underlying molecular mechanism, the proteomic signatures and responses of EGFR and downstream signals have been studied in a panel of HNSCC cell lines and tumor specimens pre- and post-gefitinib treatment.
  
  
  METHODS
  The IC(50) of gefitinib for HNSCC cell lines were determined using 3-(4,5-dmethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide proliferation assay. The effects of gefitinib on activation of EGFR and downstream signaling molecules were determined by Western blot, ELISA, and reverse-phase protein microarray (RPMA). The biomarkers involved in the signaling pathways were examined in HNSCC tumor specimens from patients in a phase I gefitinib trial.
  
  
  RESULTS
  In vitro, gefitinib inhibited cell proliferation with differing IC(50), and suppressed activation of EGFR and downstream signaling molecules protein kinase B (AKT), extracellular signal-regulated kinase 1/2, signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappaB. The drug sensitivity was statistically correlated with activation of phosphorylated AKT (p-AKT) and phosphorylated STAT3 (p-STAT3) detected by ELISA, and consistent with results measured by RPMA. In patient samples, a broad suppression of activation of EGFR and downstream signaling molecules was observed in a molecular responder patient, in contrast to a lack of inhibition or increased activation of biomarkers in different pathways in nonresponder patients.
  
  
  CONCLUSIONS
  Gefitinib sensitivity is correlated with p-AKT and p-STAT3 activation in HNSCC cell lines and tumor specimens. p-AKT and p-STAT3 could serve as potentially useful biomarkers and drug targets for further development of novel therapeutic agents for HNSCC.
authors.affiliation: Tumor Biology Section, Head and Neck Surgery Branch, National Institute on Deafness and Other Communication Disorders, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA.
authors.forename: Francisco G
authors.initials: FG
authors.lastname: Pernas
authors.forename: Clint T
authors.initials: CT
authors.lastname: Allen
authors.forename: Mary E
authors.initials: ME
authors.lastname: Winters
authors.forename: Bin
authors.initials: B
authors.lastname: Yan
authors.forename: Jay
authors.initials: J
authors.lastname: Friedman
authors.forename: Bhavana
authors.initials: B
authors.lastname: Dabir
authors.forename: Kunal
authors.initials: K
authors.lastname: Saigal
authors.forename: Gerhard S
authors.initials: GS
authors.lastname: Mundinger
authors.forename: Xiaojiang
authors.initials: X
authors.lastname: Xu
authors.forename: John C
authors.initials: JC
authors.lastname: Morris
authors.forename: Katherine R
authors.initials: KR
authors.lastname: Calvo
authors.forename: Carter
authors.initials: C
authors.lastname: Van Waes
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen