Do-Hyung Lee1, Jung Kweon Kim, Choun-Ki Joo. 1. Department of Ophthalmology, Ilsan Paik Hospital, Inje University Medical College, 2240 Dae Wha-dong, Ilsan-ku, Koyang, Kyunggyi-do 411-706, Korea. dhlee@ilsanpaik.ac.kr
Abstract
PURPOSE: This study was performed to elucidate the role of nuclear factor-kappaB (NF-kappaB) in the death of corneal epithelial cells after ultraviolet (UV) irradiation. METHODS: Simian virus 40-transfected human corneal epithelial cells (T-HCECs) were used in this study. Cell cultures were irradiated with a UVB (312 nm) source located 10 cm from the bottom of the slides for 10, 20, 30, or 40 s. Cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Translocation of NF-kappaB was examined by immunocytochemistry using anti-NF-kappaB p65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine and SN-50, specific NF-kappaB inhibitors, were used to confirm the role of NF-kappaB by pretreating samples for 30 min before UV irradiation, after which cytotoxicity and NF-kappaB translocation were evaluated. RESULTS: When T-HCECs were irradiated with UVB, translocation of NF-kappaB was observed with immunocytochemistry. These translocations peaked 2 h after UV irradiation during EMSA. When pretreated with sulfasalazine or SN-50, the translocation of NF-kappaB was blocked. Cellular death after UV irradiation was also markedly blocked by sulfasalazine. CONCLUSION: These findings suggest that NF-kappaB plays an important role in cellular death after UV irradiation. Copyright (c) 2005 S. Karger AG, Basel.
PURPOSE: This study was performed to elucidate the role of nuclear factor-kappaB (NF-kappaB) in the death of corneal epithelial cells after ultraviolet (UV) irradiation. METHODS:Simian virus 40-transfected human corneal epithelial cells (T-HCECs) were used in this study. Cell cultures were irradiated with a UVB (312 nm) source located 10 cm from the bottom of the slides for 10, 20, 30, or 40 s. Cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Translocation of NF-kappaB was examined by immunocytochemistry using anti-NF-kappaBp65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine and SN-50, specific NF-kappaB inhibitors, were used to confirm the role of NF-kappaB by pretreating samples for 30 min before UV irradiation, after which cytotoxicity and NF-kappaB translocation were evaluated. RESULTS: When T-HCECs were irradiated with UVB, translocation of NF-kappaB was observed with immunocytochemistry. These translocations peaked 2 h after UV irradiation during EMSA. When pretreated with sulfasalazine or SN-50, the translocation of NF-kappaB was blocked. Cellular death after UV irradiation was also markedly blocked by sulfasalazine. CONCLUSION: These findings suggest that NF-kappaB plays an important role in cellular death after UV irradiation. Copyright (c) 2005 S. Karger AG, Basel.
Authors: Sara I Van Acker; Bert Van den Bogerd; Michel Haagdorens; Vasiliki Siozopoulou; Sorcha Ní Dhubhghaill; Isabel Pintelon; Carina Koppen Journal: Cells Date: 2021-06-22 Impact factor: 6.600