Transcriptional profiling of IKK2/NF-κB- and p38 MAP kinase-dependent gene expression in TNF-α-stimulated primary human endothelial cells

Inflammatory stimulation of endothelial than 13 000 genes allowed definition of cells by tumor necrosis factor alpha (TNF-alpha) involves activation of nuclear factor kappaB (NF-kappaB) and p38 mitogen-activated protein (MAP) kinase signaling pathways. A reliable analysis of the gene expression program elicited by TNF-a and its assignment to distinct signaling pathways is not available. A sophisticated analysis of oligonucleoticle microarrays covering more than 13 000 genes allowed definition of the TNF-alpha-regulated endothelial gene expression profile and novel TINF-alpha-induced genes. Virtually all TNF-alpha-inducible genes were dependent on IkappaB kinase 2 (IKK2)/NF-kappaB activation, whereas a minor number was additionally modulated by p38. Furthermore, genes suppressed by IKK2/NF-kappaB were newly identified. Real-time reverse transcriptase-polymer- ase chain reaction (RT-PCR) and flow cytometry confirmed reliability of data. Thus, these results define a list of primary candidates for targeted modulation of endothelial functions during inflammation. (C) 2004 by The American Society of Hematology.