title: Expression of the α5 integrin gene in corneal epithelial cells cultured on tissue-engineered human extracellular matrices.
abstract: The integrin α5β1 plays a major role in corneal wound healing by promoting epithelial cell adhesion and migration over the fibronectin matrix secreted as a cellular response to corneal damage. Expression of α5 is induced when rabbit corneal epithelial cells (RCECs) are grown in the presence of fibronectin. Here, we examined whether α5 expression is similarly altered when RCECs or human corneal epithelial cells (HCECs) are grown on a reconstructed stromal matrix used as an underlying biomaterial. Mass spectrometry and immunofluorescence analyses revealed that the biomaterial matrix produced by culturing human corneal fibroblasts with ascorbic acid (ECM/35d) contains several types of collagens, fibronectin, tenascin and proteoglycans. Results from transfection of CAT/α5-promoter plasmids, Western blot and EMSA analyses indicated that ECM/35d significantly increase expression of α5 in HCECs as a result of alteration in the expression and DNA binding of the transcription factors NFI, Sp1, AP-1 and PAX6. The biological significance of this biomaterial substitute on the expression of the α5 gene may therefore contribute to better understand the function played by the α5β1 integrin during corneal wound healing.
authors.affiliation: CUO-Recherche, Centre de recherche FRQS du CHU de Québec, Québec, Canada.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Lake
authors.forename: Karine
authors.initials: K
authors.lastname: Zaniolo
authors.forename: Manon
authors.initials: M
authors.lastname: Gaudreault
authors.forename: Patrick
authors.initials: P
authors.lastname: Carrier
authors.forename: Alexandre
authors.initials: A
authors.lastname: Deschambault
authors.forename: Richard
authors.initials: R
authors.lastname: Bazin
authors.forename: Lucie
authors.initials: L
authors.lastname: Germain
authors.forename: Christian
authors.initials: C
authors.lastname: Salesse
authors.forename: Sylvain L
authors.initials: SL
authors.lastname: Guérin
mesh_terms: D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D039482:Integrin alpha5 / Q000235:genetics*; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D011817:Rabbits; D023822:Tissue Engineering
chemical_list: D039482:Integrin alpha5