Kruppel-like factor 6 (KLF6) affects the promoter activity of the alpha1-proteinase inhibitor gene
| Autor: | Joe¨l Sugar, Hiroshi Nakamura, Fabienne De Graeve, Hiroshi Sakai, Bernard Dastugue, Vincent Sapin, Fre´de´ric Chiambaretta, Yasuhiro Maruyama, Geoffroy Marceau, Beatrice Y.J.T. Yue, Danie`le Rigal |
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| Rok vydání: | 2006 |
| Předmět: |
Adult
Keratoconus genetic structures Adolescent Blotting Western Molecular Sequence Data Kruppel-Like Transcription Factors Down-Regulation Biology Transfection Cathepsin B Proto-Oncogene Proteins medicine Kruppel-Like Factor 6 Humans RNA Messenger Child Fluorescent Antibody Technique Indirect Promoter Regions Genetic Transcription factor Cells Cultured In Situ Hybridization Corneal epithelium Aged Aged 80 and over Expression vector Base Sequence Reverse Transcriptase Polymerase Chain Reaction Epithelium Corneal Promoter Zinc Fingers Middle Aged medicine.disease Molecular biology eye diseases Lysosomal acid phosphatase KLF6 medicine.anatomical_structure Gene Expression Regulation alpha 1-Antitrypsin sense organs |
| Zdroj: | Investigative ophthalmologyvisual science. 47(2) |
| ISSN: | 0146-0404 |
| Popis: | PURPOSE Keratoconus is a progressive disease that thins and scars the cornea. In keratoconus corneas, levels of degradative enzymes, including lysosomal acid phosphatase (LAP) and cathepsin B, are elevated, and those of inhibitors alpha1-proteinase inhibitor (alpha1-PI) and alpha2-macroglobulin (alpha2-M) are reduced. The present study explored the possible involvement in keratoconus of Kruppel-like factor 6 (KLF6), a transcription factor previously described to be essential for the integrity of the corneal epithelium. The transcript and proteins level of KLF6 and its action in regulating the genes affected in keratoconus were examined in this study. METHODS Semiquantitative RT-PCR, Western blot analysis, immunofluorescence and in situ hybridization were used to investigate the expression of KLF6 mRNA and protein in normal and keratoconus corneas. Modulation by KLF6 of the promoter activity of alpha1-PI, LAP, cathepsin B, and alpha2-M genes was studied after transient transfection of KLF6 expression plasmid into corneal epithelial cells using promoter-reporter gene assays. Chromatin immunoprecipitation (ChIP) assays were performed to confirm the interactions between KLF6 and promoters of the genes affected in keratoconus. RESULTS A global increased expression of the transcription factor KLF6 in terms of mRNAs and proteins was observed in total cornea and/or the epithelium in a substantial number of the keratoconus specimens. The promoter activity of the human alpha1-PI gene was suppressed by expression of KLF6 in corneal epithelial cells. The ChIP assay confirmed a physical interaction between KLF6 and the alpha1-PI promoter. CONCLUSIONS Transcription factor KLF6 downregulates the alpha1-PI gene in corneal epithelial cells and may thereby be involved in keratoconus. |
| Databáze: | OpenAIRE |
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