Translocation of nuclear factor-kappaB on corneal epithelial cells induced by ultraviolet B irradiation.
OBJECTIVE
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.


CONCLUSIONS
These findings suggest that NF-kappaB plays an important role in cellular death after UV irradiation.
Lee
Do-Hyung
DH
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
Kim
Jung Kweon
JK
Joo
Choun-Ki
CK
D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D002472:Cell Transformation, Viral; D024202:Electrophoretic Mobility Shift Assay; D019573:Epithelium, Corneal / Q000378:metabolism / Q000528:radiation effects*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D013539:Simian virus 40; D014178:Translocation, Genetic; D014466:Ultraviolet Rays
D016328:NF-kappa B