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Published on: January 12, 2020
NF-κB activation by peroxynitrite through IκBα-dependent phosphorylation versus nitration in colon cancer cells
Einat Gochman1, Jamal Mahajna, Abraham Z Reznick
1Department of Anatomy and Cell Biology, The Rappaport Faculty of Medicine, Technion, Haifa, Israel.
Background:
Peroxynitrite has been proposed to activate Nuclear Factor κappa beta (NF-κB) in a non-canonical or aberrant pathway and to contribute to pathology of many diseases.
Methods:
The activation of NF-κB by peroxynitrite was assessed by Western blot, immunoprecipitation, RT-PCR, and image stream analysis.
Results:
Our work showed that in HT-29 cancer cells, peroxynitrite can cause nitration of Inhibitory protein kappa B alpha (IκBα) at the expense of its phosphorylation. This led to a decrease in the degradation and re-synthesis of IκBα. Similar findings were observed for mRNA levels assessed by RT-PCR. Exposure of HT-29 cells to p38 inhibitor SB202190, prior to stimulation, resulted in a dramatic decrease of IκBα kinase and IκBα phosphorylation and caused an increase of peroxynitrite-mediated nitration of IκBα, indicating that peroxynitrite may activate NF-κB via dual mechanism of IκBα phosphorylation which is p38 dependent, as well as IκBα nitration.
Conclusion:
Our findings demonstrate a possible interaction of the p38 pathway with the NF-κB pathway under peroxynitrite stimulation.
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