Cellular death mediated by nuclear factor kappa B (NF-kappaB) translocation in cultured human lens epithelial cells after ultraviolet-B irradiation.
OBJECTIVE
To determine the role of nuclear factor kappa B (NF-kappaB) in the death of lens epithelial cells (LECs) after ultraviolet (UV) irradiation.


METHODS
Department of Ophthalmology, Ilsan Paik Hospital, Inje University, Korea.


METHODS
Cultures of simian virus 40 transfected human LECs (HLE B-3 cells) were were irradiated with a UVB source (312 nm) located 10 cm from the bottom of the slides for 1, 2, 3, or 4 minutes. Cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide method. Translocation of NF-kappaB was examined by immunocytochemistry using anti-NF-kappaB p65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine, a specific NF-kappaB inhibitor, was used to confirm the role of NF-kappaB by pretreating samples for 30 minutes before UV irradiation, after which cytotoxicity and NF-kappaB translocation were evaluated.


RESULTS
When HLE B-3 cells were irradiated with UVB, translocation of NF-kappaB was observed with immunocytochemistry. These translocations peaked during EMSA 6 hours after UV irradiation. In HLE B-3 cells pretreated with sulfasalazine, the translocation of NF-kappaB was blocked. Cellular death after UV irradiation was also markedly reduced by sulfasalazine pretreatment. Ultraviolet irradiation can translocate NF-kappaB, and sulfasalazine is a useful blocking agent in this pathway. In this experimental model, sulfasalazine prevented cellular death after UV irradiation.


CONCLUSIONS
The findings suggest that NF-kappaB plays an important role in cellular death after UV irradiation.
Lee
Do Hyung
DH
Department of Ophthalmology, Inje University, Ilsan Paik Hospital, Korea. dhlee@ilsanpaik.ac.kr
Cho
Kyung Sun
KS
Park
Sang-Gyu
SG
Kim
Eung Kwon
EK
Joo
Choun-Ki
CK
D017209:Apoptosis; D002470:Cell Survival; D002478:Cells, Cultured; D024202:Electrophoretic Mobility Shift Assay; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007908:Lens, Crystalline / Q000166:cytology*; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012460:Sulfasalazine / Q000494:pharmacology; D014466:Ultraviolet Rays
D016328:NF-kappa B; D012460:Sulfasalazine