Translocation of Nuclear Factor-κB on Corneal Epithelial Cells Induced by Ultraviolet B Irradiation

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Abstract
Purpose: This study was performed to elucidate the role of nuclear factor-ĸB (NF-ĸB) 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-ĸB was examined by immunocytochemistry using anti-NF-ĸB p65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine and SN-50, specific NF-ĸB inhibitors, were used to confirm the role of NF-ĸB by pretreating samples for 30 min before UV irradiation, after which cytotoxicity and NF-ĸB translocation were evaluated. Results: When T-HCECs were irradiated with UVB, translocation of NF-ĸB 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-ĸB was blocked. Cellular death after UV irradiation was also markedly blocked by sulfasalazine. Conclusion: These findings suggest that NF-ĸB plays an important role in cellular death after UV irradiation.