The Role of Nuclear Factor Kappa B on Lens Epithelial Cells after Ultraviolet B Irradiation.
- Author:
Do Hyung LEE
1
;
Jung Kweon KIM
;
Sae Hoon OH
;
Eung Kwon KIM
;
Choun Ki JOO
Author Information
1. Department of Ophthalmology, Ilsan Paik Hospital, College of Medicine, Inje University, Korea.
- Publication Type:Original Article
- Keywords:
Human lens epithelial cells;
Nuclear factor kappa B;
Ultraviolet irradiation
- MeSH:
Electrophoretic Mobility Shift Assay;
Epithelial Cells*;
Humans;
Immunohistochemistry;
NF-kappa B*;
Simian virus 40;
Sulfasalazine;
Transcription Factor RelA
- From:Journal of the Korean Ophthalmological Society
2004;45(3):500-506
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: This study was performed to determine the role of nuclear factor kappa B (NF-kappa B) on the lens epithelial cell death after ultraviolet (UV) irradiation. METHODS: Simian virus 40 transfected human lens epithelial cells (HLE B-3 cells) were used in this study. UVB located at 10cm from the bottom was irradiated during 1, 2, 3 and 4 minutes. To measure the cytotoxicity MTT assay was used. Translocation of NF-kappa B was examined by immunocytochemistry with anti NF-kappa B p65 antibody and electrophoretic mobility shift assay (EMSA). To confirm the role of NF-kappa B, the cells were pretreated with sulfasalazine, a specific inhibitor of NF-kappa B, for 30 minutes before irradiation, and cytotoxicity and translocation of NF-kappa B were evaluated. RESULTS: UV irradiation produced a progressive cytotoxic effect in cultured HLE B-3 cells after 1 minute and maximum cytotoxicity was reached after 3 minutes irradiation. When HLE B-3 cells were irradiated with UVB, the translocation of NF-kappa B was observed in immunocytochemistry. These translocations were peaked 6 hours after UV irradiation in EMSA. In HLE B-3 cells pretreated with sulfasalazine, the translocation of NF-kappa B was blocked. The cellular death after UV irradiation was markedly blocked by sulfasalazine. UV irradiation can translocate NF-kappa B and sulfasalazine is a useful blocking agent in this pathway. In addition, sulfasalazine can prevent cellular death after UV irradiation. CONCLUSIONS: These findings suggest that NF-kappa B plays an important role in cellular death after UV irradiation.
