| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 From the Department of Ophthalmology, University of Clermont-Ferrand, France; 2 National Institute of Health and Medical Research (INSERM) Unit 384, Faculty of Medicine, Clermont-Ferrand, France; the 3 Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio; and the 4 Department of Ophthalmology, University of Washington School of Medicine, Seattle, Washington.
PURPOSE. The keratin-12 (K12) protein is essential for the integrity of the corneal epithelium. This study was conducted to investigate the possible involvement of Krüppel-like factor 6 (KLF6) in the corneal regulation of K12 gene expression, in view of the presence of one KLF6 potential binding site in the human K12 promoter and the known role of KLF6 in regulating keratin gene expression.
METHODS. RT-PCR, Western blot analysis, and immunolocalization experiments were used to investigate the expression of KLF6 mRNA and protein in five human total corneas. The same experimental design was used to explore human corneal epithelial (HCE) cells in 20 patients and a HCE cell line. The ability of the KLF6 protein to modulate K12 promoter activity was studied in the HCE cell line, by transient transfections with a KLF6 expression plasmid and promoter-reporter gene assays. Gel-shift assays were performed to confirm the interactions between the KLF6 protein and specific sequences of the K12 promoter.
RESULTS. The presence of KLF6 transcripts and proteins in human total corneal extracts was demonstrated. Immunohistofluorescence experiments showed positive staining specifically present in the corneal epithelial layer. KLF6 transcripts and proteins were also present in corneal epithelial cells in 20 patients and the HCE cell line. Transient transfections of KLF6 showed statistical transactivation of the K12 promoter in HCE cells. The gel-shift assay showed a physical interaction between KLF6 and the K12 promoter.
CONCLUSIONS. The expression of KLF6 in HCE cells and its role in the regulation of K12 gene expression were demonstrated.
This article has been cited by other articles:
|
|
 |
|
  S. K. Swamynathan, J. P. Katz, K. H. Kaestner, R. Ashery-Padan, M. A. Crawford, and J. Piatigorsky Conditional Deletion of the Mouse Klf4 Gene Results in Corneal Epithelial Fragility, Stromal Edema, and Loss of Conjunctival Goblet Cells Mol. Cell. Biol., January 1, 2007; 27(1): 182 - 194. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Chiambaretta, H. Nakamura, F. De Graeve, H. Sakai, G. Marceau, Y. Maruyama, D. Rigal, B. Dastugue, J. Sugar, B. Y. J. T. Yue, et al. Kruppel-like Factor 6 (KLF6) Affects the Promoter Activity of the {alpha}1-Proteinase Inhibitor Gene Invest. Ophthalmol. Vis. Sci., February 1, 2006; 47(2): 582 - 590. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Nakamura, F. Chiambaretta, J. Sugar, V. Sapin, and B. Y. J. T. Yue Developmentally Regulated Expression of KLF6 in the Mouse Cornea and Lens Invest. Ophthalmol. Vis. Sci., December 1, 2004; 45(12): 4327 - 4332. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Ito, M. Uchiyama, M. Kondo, S. Mori, N. Usami, O. Maeda, T. Kawabe, Y. Hasegawa, K. Shimokata, and Y. Sekido Kruppel-Like Factor 6 Is Frequently Down-Regulated and Induces Apoptosis in Non-Small Cell Lung Cancer Cells Cancer Res., June 1, 2004; 64(11): 3838 - 3843. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
