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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
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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.
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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.
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Contributor(s) |
Lake J, Zaniolo K, Gingras M, Salesse C, Guérin SL |
Citation missing |
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Submission date |
Jan 29, 2015 |
Last update date |
Nov 27, 2018 |
Contact name |
Karine Zaniolo |
E-mail(s) |
karine.zaniolo@gmail.com
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Organization name |
Centre de recherche du CHU de Québec
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Department |
CUO-Recherche
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Lab |
Sylvain Guérin
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Street address |
1050 Chemin Sainte-Foy
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City |
Québec |
State/province |
Québec |
ZIP/Postal code |
G1S 4L8 |
Country |
Canada |
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Platforms (1) |
GPL13607 |
Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Feature Number version) |
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Samples (6)
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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 |
GSM1595871 |
Primary human corneal epithelial cells Collagen type I rep2 |
GSM1595872 |
Primary human corneal epithelial cells Collagen type IV rep1 |
GSM1595873 |
Primary human corneal epithelial cells Collagen type IV rep2 |
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Relations |
BioProject |
PRJNA273984 |