title: Identification of biomarkers in human head and neck tumor cell lines that predict for in vitro sensitivity to gefitinib.
abstract: Potential biomarkers were identified for in vitro sensitivity to the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor gefitinib in head and neck cancer. Gefitinib sensitivity was determined in cell lines, followed by transcript profiling coupled with a novel pathway analysis approach. Eleven cell lines were highly sensitive to gefitinib (inhibitor concentration required to give 50% growth inhibition [GI(50)] < 1 microM), three had intermediate sensitivity (GI(50) 1-7 microM), and six were resistant (GI(50) > 7 microM); an exploratory principal component analysis revealed a separation between the genomic profiles of sensitive and resistant cell lines. Subsequently, a hypothesis-driven analysis of Affymetrix data (Affymetrix, Inc., Santa Clara, CA, USA) revealed higher mRNA levels for E-cadherin (CDH1); transforming growth factor, alpha (TGF-alpha); amphiregulin (AREG); FLJ22662; EGFR; p21-activated kinase 6 (PAK6); glutathione S-transferase Pi (GSTP1); and ATP-binding cassette, subfamily C, member 5 (ABCC5) in sensitive versus resistant cell lines. A hypothesis-free analysis identified 46 gene transcripts that were strongly differentiated, seven of which had a known association with EGFR and head and neck cancer (human EGF receptor 3 [HER3], TGF-alpha, CDH1, EGFR, keratin 16 [KRT16], fibroblast growth factor 2 [FGF2], and cortactin [CTTN]). Polymerase chain reaction (PCR) and enzyme-linked immunoabsorbant assay analysis confirmed Affymetrix data, and EGFR gene mutation, amplification, and genomic gain correlated strongly with gefitinib sensitivity. We identified biomarkers that predict for in vitro responsiveness to gefitinib, seven of which have known association with EGFR and head and neck cancer. These in vitro predictive biomarkers may have potential utility in the clinic and warrant further investigation.
authors.affiliation: Fomerly of AstraZeneca, Alderly Park, Macclesfield, UK.
authors.forename: D Mark
authors.initials: DM
authors.lastname: Hickinson
authors.forename: Gayle B
authors.initials: GB
authors.lastname: Marshall
authors.forename: Garry J
authors.initials: GJ
authors.lastname: Beran
authors.forename: Marileila
authors.initials: M
authors.lastname: Varella-Garcia
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Mills
authors.forename: Marie C
authors.initials: MC
authors.lastname: South
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Cassidy
authors.forename: Kerry L
authors.initials: KL
authors.lastname: Acheson
authors.forename: Gael
authors.initials: G
authors.lastname: McWalter
authors.forename: Rose M
authors.initials: RM
authors.lastname: McCormack
authors.forename: Paul A
authors.initials: PA
authors.lastname: Bunn
authors.forename: Tim
authors.initials: T
authors.lastname: French
authors.forename: Alex
authors.initials: A
authors.lastname: Graham
authors.forename: Brian R
authors.initials: BR
authors.lastname: Holloway
authors.forename: Fred R
authors.initials: FR
authors.lastname: Hirsch
authors.forename: Georgina
authors.initials: G
authors.lastname: Speake
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D019008:Drug Resistance, Neoplasm / Q000187:drug effects* / Q000235:genetics; D004354:Drug Screening Assays, Antitumor; D004797:Enzyme-Linked Immunosorbent Assay; D000077156:Gefitinib; D018628:Gene Dosage / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D015894:Genome, Human / Q000235:genetics; D006258:Head and Neck Neoplasms / Q000235:genetics*; D006801:Humans; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D011799:Quinazolines / Q000494:pharmacology* / Q000627:therapeutic use; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D011799:Quinazolines; D012333:RNA, Messenger; D000077156:Gefitinib