A role for c-Raf kinase and Ha-Ras in cytokine-mediated induction of cell adhesion molecules.

Cytokines such as tumor necrosis factor (TNFalpha) play fundamental roles in the pathophysiology of inflammation and immunity-related diseases. Despite rapid advances in our understanding of cytokine biology in recent years, definitive knowledge of the cytokine cell signaling pathways remains elusive due to the enormous complexity of these pathways and the lack of specific biological tools and reagents. Using highly specific antisense oligonucleotides that target the mRNA encoding c-Raf kinase and Ha-Ras, we show here that inhibition of c-raf and Ha-ras expression blocks the up-regulation of E-selectin and vascular adhesion molecule-1 induced by TNFalpha in endothelial cells. Induction of intercellular adhesion molecule-1 was also reduced, although to a much lesser extent, by treatment with antisense oligonucleotides. We also show that inhibition of c-raf kinase expression decreased extracellular signal-regulated kinase and c-Jun N-terminal kinase (JNK) kinase stimulation by TNFalpha. Furthermore, antisense inhibition of JNK2 also blocked TNFalpha-mediated induction of E-selectin, whereas PD98059 (mitogen-activated protein kinase kinase 1 and 2 inhibitor) had no effect on this process. These results indicate that TNFalpha induction of E-selectin and vascular adhesion molecule-1 in endothelial cells occurs through signaling pathways that are, at least in part, dependent on c-Raf kinase, Ha-Ras, and JNK2.