title: Transcriptional profiling identifies cyclin D1 as a critical downstream effector of mutant epidermal growth factor receptor signaling.
abstract: Activating mutations in the epidermal growth factor receptor (EGFR) tyrosine kinase domain determine responsiveness to EGFR tyrosine kinase inhibitors in patients with advanced non-small cell lung cancer (NSCLC). The modulation of transcriptional pathways by mutant EGFR signaling is not fully understood. Previously, we and others identified a single base pair change leading to a threonine to methionine (T790M) amino acid alteration in the ATP-binding pocket of the EGFR as a common mechanism of acquired resistance. The gefitinib-resistant, T790M-mutant H1975 NSCLC cell line undergoes prominent growth arrest and apoptosis when treated with the irreversible EGFR inhibitor, CL-387,785. We did a transcriptional profiling study of mutant EGFR target genes that are differentially expressed in the "resistant" gefitinib-treated and the "sensitive" CL387,785-treated H1975 cells to identify the pivotal transcriptional changes in NSCLC with EGFR-activating mutations. We identified a small subset of early gene changes, including significant reduction of cyclin D1 as a result of EGFR inhibition by CL-387,785 but not by gefitinib. The reduction in cyclin D1 transcription was associated with subsequent suppression of E2F-responsive genes, consistent with proliferation arrest. Furthermore, cyclin D1 expression was higher in EGFR-mutant lung cancer cells compared with cells with wild-type EGFR. EGFR-mutant cells were routinely sensitive to the cyclin-dependent kinase inhibitor flavopiridol, confirming the functional relevance of the cyclin D axis. These studies suggest that cyclin D1 may contribute to the emergence of EGFR-driven tumorigenesis and can be an alternative target of therapy.
authors.affiliation: Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Susumu
authors.initials: S
authors.lastname: Kobayashi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shimamura
authors.forename: Stefano
authors.initials: S
authors.lastname: Monti
authors.forename: Ulrich
authors.initials: U
authors.lastname: Steidl
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hetherington
authors.forename: April M
authors.initials: AM
authors.lastname: Lowell
authors.forename: Todd
authors.initials: T
authors.lastname: Golub
authors.forename: Matthew
authors.initials: M
authors.lastname: Meyerson
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Tenen
authors.forename: Geoffrey I
authors.initials: GI
authors.lastname: Shapiro
authors.forename: Balázs
authors.initials: B
authors.lastname: Halmos
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D015153:Blotting, Western; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D056741:Cyclin D; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors; D016213:Cyclins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D000069347:Erlotinib Hydrochloride; D005419:Flavonoids / Q000494:pharmacology; D000077156:Gefitinib; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D020125:Mutation, Missense / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010880:Piperidines / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics; D014162:Transfection
chemical_list: D000970:Antineoplastic Agents; D056741:Cyclin D; D016213:Cyclins; D005419:Flavonoids; D050505:Mutant Proteins; D010880:Piperidines; D011799:Quinazolines; C077990:alvocidib; C118029:CL 387785; D000069347:Erlotinib Hydrochloride; D066246:ErbB Receptors; D018844:Cyclin-Dependent Kinases; D000077156:Gefitinib