Haoxiang Xu1, Yan Yan, Li Li, Shiguang Peng, Tao Qu, Baoxi Wang. 1. Department of Dermatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Abstract
BACKGROUND: After irradiation with a high dose of ultraviolet B (UVB), cells undergo apoptosis. Caspase-8 and -3 are key mediators of apoptosis in many cells. Vimentin, an important cytoskeleton component, can be cleaved by caspase-3, -6, -7 and -8. Cell apoptosis is promoted via caspase-triggered proteolysis of vimentin. In this study, we explored the roles of caspase-8 and -3 and the changes in vimentin expression in UVB-induced apoptosis of human dermal fibroblasts. METHODS: Skin fibroblasts were irradiated with 150 mJ/cm(2) UVB and cell death was monitored by the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and Hoechst staining. Caspase-8 and -3 activities were detected by the caspase activity assay. Vimentin expression was assessed by immunofluorescence and Western blot. RESULTS: Caspase-8 and -3 were activated by 150 mJ/cm(2) UVB irradiation. Caspase-8 and -3 activities changed in a time-dependent way after UVB irradiation to induce apoptosis of fibroblasts, and caspase-8 and -3 interacted with each other in this process. However, their substrate, vimentin, showed an enhanced expression over time after UVB irradiation. CONCLUSIONS: UVB-triggered apoptosis of fibroblasts was dependent on the activation of caspase-8 and -3 with an increased expression of vimentin.
BACKGROUND: After irradiation with a high dose of ultraviolet B (UVB), cells undergo apoptosis. Caspase-8 and -3 are key mediators of apoptosis in many cells. Vimentin, an important cytoskeleton component, can be cleaved by caspase-3, -6, -7 and -8. Cell apoptosis is promoted via caspase-triggered proteolysis of vimentin. In this study, we explored the roles of caspase-8 and -3 and the changes in vimentin expression in UVB-induced apoptosis of human dermal fibroblasts. METHODS: Skin fibroblasts were irradiated with 150 mJ/cm(2) UVB and cell death was monitored by the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and Hoechst staining. Caspase-8 and -3 activities were detected by the caspase activity assay. Vimentin expression was assessed by immunofluorescence and Western blot. RESULTS:Caspase-8 and -3 were activated by 150 mJ/cm(2) UVB irradiation. Caspase-8 and -3 activities changed in a time-dependent way after UVB irradiation to induce apoptosis of fibroblasts, and caspase-8 and -3 interacted with each other in this process. However, their substrate, vimentin, showed an enhanced expression over time after UVB irradiation. CONCLUSIONS: UVB-triggered apoptosis of fibroblasts was dependent on the activation of caspase-8 and -3 with an increased expression of vimentin.
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