- Authors: Janus, P., Szoltysek, K., Zajac, G., Stokowy, T., Walaszczyk, A., Widlak, W., Wojtas, B., Gielniewski, B., Iwanaszko, M., Braun, R., Cockell, S., Perkins, N. D., Kimmel, M., Widlak, P.
- Year: 2018
- Journal: Cell Signal 46 23-31
- Applications: in vitro / siRNA / INTERFERin
- Cell type: U-2 OS
Description: Human bone osteosarcoma
Known as: U2OS
The NF-kappaB transcription factors are activated via diverse molecular mechanisms in response to various types of stimuli. A plethora of functions associated with specific sets of target genes could be regulated differentially by this factor, affecting cellular response to stress including an anticancer treatment. Here we aimed to compare subsets of NF-kappaB-dependent genes induced in cells stimulated with a pro-inflammatory cytokine and in cells damaged by a high dose of ionizing radiation (4 and 10Gy). The RelA-containing NF-kappaB species were activated by the canonical TNFalpha-induced and the atypical radiation-induced pathways in human osteosarcoma cells. NF-kappaB-dependent genes were identified using the gene expression profiling (by RNA-Seq) in cells with downregulated RELA combined with the global profiling of RelA binding sites (by ChIP-Seq), with subsequent validation of selected candidates by quantitative PCR. There were 37 NF-kappaB-dependent protein-coding genes identified: in all cases RelA bound in their regulatory regions upon activation while downregulation of RELA suppressed their stimulus-induced upregulation, which apparently indicated the positive regulation mode. This set of genes included a few "novel" NF-kappaB-dependent species. Moreover, the evidence for possible negative regulation of ATF3 gene by NF-kappaB was collected. The kinetics of the NF-kappaB activation was slower in cells exposed to radiation than in cytokine-stimulated ones. However, subsets of NF-kappaB-dependent genes upregulated by both types of stimuli were essentially the same. Hence, one should expect that similar cellular processes resulting from activation of the NF-kappaB pathway could be induced in cells responding to pro-inflammatory cytokines and in cells where so-called "sterile inflammation" response was initiated by radiation-induced damage.