Document Detail


Imaging of nuclear factor κB activation induced by ionizing radiation in human embryonic kidney (HEK) cells.
MedLine Citation:
PMID:  24880906     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Ionizing radiation modulates several signaling pathways resulting in transcription factor activation. Nuclear factor kappa B (NF-κB) is one of the most important transcription factors that respond to changes in the environment of a mammalian cell. NF-κB plays a key role not only in inflammation and immune regulation but also in cellular radiation response. In response to DNA damage, NF-κB might inhibit apoptosis and promote carcinogenesis. Our previous studies showed that ionizing radiation is very effective in inducing biological damages. Therefore, it is important to understand the radiation-induced NF-κB signaling cascade. The current study aims to improve existing mammalian cell-based reporter assays for NF-κB activation by the use of DD-tdTomato which is a destabilized variant of red fluorescent protein tdTomato. It is demonstrated that exposure of recombinant human embryonic kidney cells (HEK/293 transfected with a reporter constructs containing NF-κB binding sites in its promoter) to ionizing radiation induces NF-κB-dependent DD-tdTomato expression. Using this reporter assays, NF-κB signaling in mammalian cells was monitored by flow cytometry and fluorescence microscopy. Activation of NF-κB by the canonical pathway was found to be quicker than by the genotoxin- and stress-induced pathway. X-rays activate NF-κB in HEK cells in a dose-dependent manner, and the extent of NF-κB activation is higher as compared to camptothecin.
Authors:
Arif Ali Chishti; Christa Baumstark-Khan; Christine E Hellweg; Günther Reitz
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-6-1
Journal Detail:
Title:  Radiation and environmental biophysics     Volume:  -     ISSN:  1432-2099     ISO Abbreviation:  Radiat Environ Biophys     Publication Date:  2014 Jun 
Date Detail:
Created Date:  2014-6-1     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0415677     Medline TA:  Radiat Environ Biophys     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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