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Series GSE65421 Query DataSets for GSE65421
Status Public on Jan 30, 2015
Title Functional impact of collagens on the activity directed by the promoter of the a5 integrin subunit gene in corneal epithelial cells
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Important remodeling of the extracellular matrix is a hallmark of corneal wound healing. Besides the massive secretion of fibronectin (FN) that characterizes the early step of that process, alterations in the secretion of collagens also occurs as part of this wound healingresponse. In this study, we examined whether expression of the gene encoding the α5 subunit from the FN binding integrin α5β1 changes as corneal epithelial cells (CECs) are cultured in the presence of collagens. Responsiveness of the α5 gene toward collagen was determined by transfection of recombinant plasmids bearing the CAT reporter gene under the control of various segments from the human α5 promoter into primary cultured rabbit (RCECs) and human (HCECs) CECs cultured on BSA or on collagens. Electrophoretic mobility shift assays (EMSAs) and Western blot analyses were used to monitor both the DNA binding and expression of the transcription factors required to ensure basal transcription of the α5 gene. The influence collagens on the patterns of genes expressed by HCECs was also studied be gene profiling on microarrays. All collagen types repressed the transcriptional activity directed by the full-length α5 promoter/CAT construct in confluent CECs. A moderate increase in α5 promoter activity was observed in subconfluent RCECs grown on CIV but not on CI. These collagen-dependent regulatory influences also correlated with alterations in either the expression or the DNA binding of the transcription factors Sp1/Sp3, NFI and AP-1 that ensure basal transcription of the α5 gene. Gene profiling on microarray revealed that CI more profoundly alter the pattern of genes expressed by HCECs than what is observed with CIV. Collagens considerably suppressed expression of the α5 gene in CECs at confluence suggesting that the sustained staining for CI and CIV after corneal injury may play a role in controlling adhesion to the temporary FN matrix and further differentiation of CECs into suprabasal epithelial cells through a mechanism involving the α5β1 integrin.
Overall design Primary cultures of human corneal epithelial cells cultivated on BSA (number of replicates: 2), Collagen type I (number of replicates: 2) and Collagen type IV (number of replicates: 2) matrix.
Contributor(s) Lake J, Zaniolo K, Gingras M, Salesse C, Guérin SL
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Submission date Jan 29, 2015
Last update date Nov 27, 2018
Contact name Karine Zaniolo
Organization name Centre de recherche du CHU de Québec
Department CUO-Recherche
Lab Sylvain Guérin
Street address 1050 Chemin Sainte-Foy
City Québec
State/province Québec
ZIP/Postal code G1S 4L8
Country Canada
Platforms (1)
GPL13607 Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Feature Number version)
Samples (6)
GSM1595868 Primary human corneal epithelial cells BSA rep1
GSM1595869 Primary human corneal epithelial cells BSA rep2
GSM1595870 Primary human corneal epithelial cells Collagen type I rep1
BioProject PRJNA273984

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Supplementary file Size Download File type/resource
GSE65421_RAW.tar 79.1 Mb (http)(custom) TAR (of TXT)
Processed data included within Sample table

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