Activation of NF-κB and apoptosis of intestinal epithelial cells induced by hydrogen peroxide

Objective: In vitro model of hydrogen peroxide induced apoptosis of SW-480 cells was used to investigate the role of NF-κB in the pathogenesis of reactive oxygen species induced apoptosis of intestinal epithelial cells. Methods: Ultra-structural changes were observed. Apoptosis of SW-480 cell line was determined by Annexin-V and PI double-stained flow cytometry. Nuclear translocation of NF-κB was determined by anti-NF-κB polyclonal antibody and EB double-staining. NF-κB activity was studied by electrophoretic mobility shift assays. RT-PCR was performed to study expression of NF-κB mRNA. Results: Hydrogen peroxide led to apoptosis of SW-480 cells, condensed or semilunar chromatin even apoptotic bodies could be observed. Nuclear translocation of NF-κB, increase of NF-κB activity and expression of NF-κB mRNA were found simultaneously. Conclusions: Early activation of NF-κ B may be oneof the mechanisms of apoptosis in intestinal epithelial cells by reactive oxygen species.