title: Cellular death mediated by nuclear factor kappa B (NF-kappaB) translocation in cultured human lens epithelial cells after ultraviolet-B irradiation.
abstract: 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.
authors.affiliation: Department of Ophthalmology, Inje University, Ilsan Paik Hospital, Korea. dhlee@ilsanpaik.ac.kr
authors.forename: Do Hyung
authors.initials: DH
authors.lastname: Lee
authors.forename: Kyung Sun
authors.initials: KS
authors.lastname: Cho
authors.forename: Sang-Gyu
authors.initials: SG
authors.lastname: Park
authors.forename: Eung Kwon
authors.initials: EK
authors.lastname: Kim
authors.forename: Choun-Ki
authors.initials: CK
authors.lastname: Joo
mesh_terms: 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
chemical_list: D016328:NF-kappa B; D012460:Sulfasalazine