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

Abstract

Abstract Inflammatory stimulation of endothelial cells by tumor necrosis factor α (TNF-α) involves activation of nuclear factor κB (NF-κB) and p38 mitogen-activated protein (MAP) kinase signaling pathways. A reliable analysis of the gene expression program elicited by TNF-α and its assignment to distinct signaling pathways is not available. A sophisticated analysis of oligonucleotide microarrays covering more than 13 000 genes allowed definition of the TNF-α-regulated endothelial gene expression profile and novel TNF-α-induced genes. Virtually all TNF-α-inducible genes were dependent on IκB kinase 2 (IKK2)/NF-κB activation, whereas a minor number was additionally modulated by p38. Furthermore, genes suppressed by IKK2/NF-κB were newly identified. Real-time reverse transcriptase-polymerase 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. (Blood. 2004;103:3365-3373)