Mechanisms of Apoptosis on Human Lens Epithelium after Ultraviolet Light Exposure
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Purpose: The purpose of this study is to understand the mechanism of apoptosis occurring on a cultured human lens epithelial cell line after exposure to ultraviolet (UV) light. We intended to confirm the presence of cellular tox-icity and apoptosis and to reveal the roles of p53, caspase 3 and NOXA in these processes. Methods: Cells were irradiated with an ultraviolet lamp. Cellular toxicity was measured by a 3-(4,5-dimethylth-iazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Hoechst staining and fluorescent anti-caspase 3 antibodies were used for apoptosis investigation. The quantities of p53, caspase 3, and NOXA were measured by Western blotting for to investigate the apoptosis pathway. Results: Cellular toxicity on the human lens epithelium markedly increased with time after UV exposure. On Hoechst staining, we found that apoptosis also remarkably increased after exposure to ultraviolet light, compared with a control group. In the immunochemical study using anti-caspase 3 antibodies, active caspase 3 significantly in-creased after exposure to ultraviolet light. On Western blotting, p53 decreased, while caspase 3 and NOXA increased. Conclusions: Exposure of cultured human lens epithelial cell lines to ultraviolet light induces apoptosis, which pro-motes the expression of NOXA and caspase 3 increases without increasing p53. This may suggest that UV in-duced apoptosis is caused by a p53-independent pathway in human lens epithelial cells.
Abstract
Purpose: The purpose of this study is to understand the mechanism of apoptosis occurring on a cultured human lens epithelial cell line after exposure to ultraviolet (UV) light. We intended to confirm the presence of cellular tox-icity and apoptosis and to reveal the roles of p53, caspase 3 and NOXA in these processes. Methods: Cells were irradiated with an ultraviolet lamp. Cellular toxicity was measured by a 3-(4,5-dimethylth-iazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Hoechst staining and fluorescent anti-caspase 3 antibodies were used for apoptosis investigation. The quantities of p53, caspase 3, and NOXA were measured by Western blotting for to investigate the apoptosis pathway. Results: Cellular toxicity on the human lens epithelium markedly increased with time after UV exposure. On Hoechst staining, we found that apoptosis also remarkably increased after exposure to ultraviolet light, compared with a control group. In the immunochemical study using anti-caspase 3 antibodies, active caspase 3 significantly in-creased after exposure to ultraviolet light. On Western blotting, p53 decreased, while caspase 3 and NOXA increased. Conclusions: Exposure of cultured human lens epithelial cell lines to ultraviolet light induces apoptosis, which pro-motes the expression of NOXA and caspase 3 increases without increasing p53. This may suggest that UV in-duced apoptosis is caused by a p53-independent pathway in human lens epithelial cells.
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