Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling by array of human wild type and gefitinib-resistant A431 cancer cells


ABSTRACT: A431 wild-type (wt) cancer cell line is sensitive to treatment with EGFR tyrosine kinase inhibitors (TKIs). By culturing it chronically under gefitinib, it eventually becomes resistant (A431_GR cell). We know of a few proteins involved in this mechanism of drug resistance, but a cDNA exprssion array would add information to other genes that might be involved in this resistance mechanism. We used microarrays to identify the differences in the global gene expression between A431_wt (wild-type) cells and A431_GR cells. Experiment Overall Design: Total RNA was isolated from parental (wild-type) and GR (gefitinib-resistant) A431 cells using TRIzol reagent (Invitrogen, Carlsbad, CA), followed by RNeasy Mini Kit column purification (Qiagen, Valencia, CA) including an in column DNAse clean-up using RNAse-free DNAse Set (Qiagen). Synthesis of cRNA target, its hybridization to Human Genome U133 Plus 2.0 microarrays and scanning of those arrays was performed using Affymetrix GeneChip products and reagents in accordance with the manufacturer’s recommendations at the Vanderbilt Microarray Shared Resource.

ORGANISM(S): Homo sapiens

SUBMITTER: Shizhen Emily Wang 

PROVIDER: E-GEOD-10696 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Acquired resistance to EGFR tyrosine kinase inhibitors in cancer cells is mediated by loss of IGF-binding proteins.

Guix Marta M   Faber Anthony C AC   Wang Shizhen Emily SE   Olivares Maria Graciela MG   Song Youngchul Y   Qu Sherman S   Rinehart Cammie C   Seidel Brenda B   Yee Douglas D   Arteaga Carlos L CL   Engelman Jeffrey A JA  

The Journal of clinical investigation 20080701 7


Although some cancers are initially sensitive to EGFR tyrosine kinase inhibitors (TKIs), resistance invariably develops. We investigated mechanisms of acquired resistance to the EGFR TKI gefitinib by generating gefitinib-resistant (GR) A431 squamous cancer cells. In GR cells, gefitinib reduced phosphorylation of EGFR, ErbB-3, and Erk but not Akt. These cells also showed hyperphosphorylation of the IGFI receptor (IGFIR) and constitutive association of IRS-1 with PI3K. Inhibition of IGFIR signalin  ...[more]

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