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Gene expression profile of human lung cell line (A549) treated with 2mM 59cobalt chloride during 30mn, 2h, 4h and 24h

ABSTRACT: The purpose of this work was to identify the potential signatures of cobalt exposure using a toxicogenomic approach. A time course transcriptome analysis was performed on human type II epithelial cell line A549. Cells were exposed to cobalt at midlog phase ; a medium without FCS containing cobalt (CoCl2, Sigma) or not was added for 30mn, 2h, 4h or 24h. Total RNA was isolated using the Quiagen RNeasy miniprep kit, and then labelled using the FairPlay Microarray Labelling Kit (Stratagene). Amino reactive Cy3- and Cy5- dyes (CyTMDye Post-Labelling Reactive Dye Pack, Amersham) were further chemically coupled to test and control cDNA respectively. After hybridization, microarrays were scanned with GenePix 4000B (Axon Instrument Inc., Forster City, CA). Cy3 and Cy5 fluorescence intensities for the spots were quantified after local subtraction of background noise using Genepix Pro 4.0 software (Axon Instrument Inc.) Keywords: Time course

ORGANISM(S): Homo sapiens

PROVIDER: GSE5892 | GEO | 2006/09/22

SECONDARY ACCESSION(S): PRJNA97319

REPOSITORIES: GEO

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Gene expression profile of human lung cell line (A549) treated with 2mM 59cobalt chloride during 30mn, 2h, 4h and 24h

Project description:The purpose of this work was to identify the potential signatures of cobalt exposure using a toxicogenomic approach. A time course transcriptome analysis was performed on human type II epithelial cell line A549. Cells were exposed to cobalt at midlog phase ; a medium without FCS containing cobalt (CoCl2, Sigma) or not was added for 30mn, 2h, 4h or 24h. Total RNA was isolated using the Quiagen RNeasy miniprep kit, and then labelled using the FairPlay Microarray Labelling Kit (Stratagene). Amino reactive Cy3- and Cy5- dyes (CyTMDye Post-Labelling Reactive Dye Pack, Amersham) were further chemically coupled to test and control cDNA respectively. After hybridization, microarrays were scanned with GenePix 4000B (Axon Instrument Inc., Forster City, CA). Cy3 and Cy5 fluorescence intensities for the spots were quantified after local subtraction of background noise using Genepix Pro 4.0 software (Axon Instrument Inc.) 30 mn : exposure 1 (2 microarrays) - exposure 2 (4 microarrays) 2h : exposure 1 (2 microarrays) - exposure 2 (4 microarrays) 4h : exposure 1 (2 microarrays) - exposure 2 (2 microarrays) 24h: exposure 1 (4 microarrays) - exposure 2 (6 microarrays) - exposure 3 (4 microarrays)

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