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Series GSE5450 Query DataSets for GSE5450
Status Public on Aug 04, 2006
Title Alveolar Epithelial Cell Injury with EBV Upregulates TGF-beta1 Expression
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Idiopathic pulmonary fibrosis (IPF) is a refractory and lethal interstitial lung disease characterized by alveolar epithelial cells apoptosis, fibroblast proliferation and extra-cellular matrix proteins deposition. Epstein - Barr virus (EBV) has previously been localised to alveolar epithelial cells of IPF patients. In this study we utilised a microarray based differential gene expression analysis strategy to identify potential molecular drivers of EBV associated lung fibrosis. We employed an alveolar epithelial cell line infected with EBV (A-Akata). Lytic phase infection induced in the A-Akata cells by TPA/BA treatment resulted in increase of TGFbeta1 and TIEG1 mRNA expression. Treatment of the A-Akata cells with ganciclovir,
resulted in a down regulation of both TIEG1 and TGFbeta1 expression, accompanied by a suppression of the EBV early response genes, Rta and Zta. This suppression of cell turnover mediators was correlated with an increase in cell activity index. To identify a possible role for Rta in driving apoptotic gene expression, we inhibited the Rta gene expression by silencing RNA, resulting in a decrease in TGFbeta1 and TIEG1 expression. This study identifies an apoptotic role of the EBV early response genes, as enhancer factors of TIEG1 and TGFbeta1 in EBV infected alveolar epithelial cells, potentially providing a possible mechanism for the role of EBV infection in pulmonary fibrosis.
Keywords: EBV lytic phase infection, epithelial cell apoptosis, oligonucleotide array analysis
Overall design RNA isolation, cDNA synthesis, in vitro transcription and microarray analysis of A549, EBV infected A549 (AAKata) and A549 +TGFbeta1 10ng/ml 4hours were performed as previously reported. All analysis were microarrayed in duplicate. Image files were obtained through Affymetrix GeneChip software (MAS5). Subsequently robust multichip analysis (RMA) was performed. Expression data was compared to control by ANOVA analysis, p<0.05 correlated values and a signal log ratio of 0.6 or greater (equivalent to a fold change in expression of 1.5 or greater) were taken to identify significant differential regulation. All the SLRs data resulting from the comparative analyses were reported in a graph to determine the reliability of the assay and the linearity by r2. For all the microarray assays r2 value was higher than 0.98. Using gene expression values normalised by RMA, Average Linkage Hierarchical Cluster Analysis was performed and the results visualized by TreeView software.
Contributor(s) Malizia AP, Keating DT, Smith S, Walls D, Wood AE, Doran PP, Egan JJ
Citation(s) 18621908
Submission date Aug 03, 2006
Last update date Aug 10, 2018
Contact name Andrea Patricelli Malizia
Phone 0035317166381
Fax 0035317166309
Organization name University College Dublin
Lab Genome Resource Unit
Street address 21, Nelson Street
City Dublin
ZIP/Postal code 7
Country Ireland
Platforms (1)
GPL96 [HG-U133A] Affymetrix Human Genome U133A Array
Samples (6)
GSM124905 A549_rep1
GSM124906 A549_rep2
GSM124907 AAKata_rep1
BioProject PRJNA95947

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Supplementary file Size Download File type/resource
GSE5450_RAW.tar 19.7 Mb (http)(custom) TAR (of CEL)

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