T Bourcier1, P Forgez, V Borderie, S Scheer, W Rostène, L Laroche. 1. Cornea Bank, AP-HP, Paris VI University, the. Institut National de la Santé et de la Recherche Médicale (U33g), Saint-Antoine Hospital, Paris, France. bourcier@quinze-vingts.fr
Abstract
PURPOSE: To investigate whether human corneal epithelial cells express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells. METHODS: Human corneal epithelial cells were cultured in medium supplemented with various concentrations of DEX (ranging from 10(-10) to 10(-4) M). Cell proliferation was analyzed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfop henyl) -2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Apoptosis was studied by nucleus labeling using a fluorescent dye and immunostaining by APO 2.7 at 6 days of culture. GR mRNA was detected in corneal epithelium and cultured corneal epithelial cells by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the epithelial cells was performed with a monoclonal anti-human GR. RESULTS: RT-PCR and immunocytochemistry showed the expression of GR (mRNA and protein) in corneal epithelial cells. DEX significantly increased corneal epithelial cell proliferation with concentrations ranging from 10(-10) to 10(-6) M, with a maximum effect at 10(-7) M (P < 0.005). However, DEX also induced apoptosis of cultured corneal epithelial cells at any concentration used. CONCLUSIONS: These results indicate that human corneal epithelial cells express the GR and proliferate in response to DEX stimulation which also induces corneal epithelial cell apoptosis.
PURPOSE: To investigate whether human corneal epithelial cells express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells. METHODS:Human corneal epithelial cells were cultured in medium supplemented with various concentrations of DEX (ranging from 10(-10) to 10(-4) M). Cell proliferation was analyzed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfop henyl) -2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Apoptosis was studied by nucleus labeling using a fluorescent dye and immunostaining by APO 2.7 at 6 days of culture. GR mRNA was detected in corneal epithelium and cultured corneal epithelial cells by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the epithelial cells was performed with a monoclonal anti-humanGR. RESULTS: RT-PCR and immunocytochemistry showed the expression of GR (mRNA and protein) in corneal epithelial cells. DEX significantly increased corneal epithelial cell proliferation with concentrations ranging from 10(-10) to 10(-6) M, with a maximum effect at 10(-7) M (P < 0.005). However, DEX also induced apoptosis of cultured corneal epithelial cells at any concentration used. CONCLUSIONS: These results indicate that human corneal epithelial cells express the GR and proliferate in response to DEX stimulation which also induces corneal epithelial cell apoptosis.
Authors: M Elizabeth Hartnett; David J Martiniuk; Yuta Saito; Pete Geisen; Lynda J Peterson; Janet R McColm Journal: Invest Ophthalmol Vis Sci Date: 2006-11 Impact factor: 4.799