Inhibition of mammalian target of rapamycin potentiates thrombin-induced intercellular adhesion molecule-1 expression by accelerating and stabilizing NF-κB activation in endothelial cells

We addressed the regulatory function of mammalian target of rapamycin (mTOR) in the mechanism of thrombin-induced ICAM-1 gene expression in endothelial cells. Pretreatment of HUVECs with rapamycin, an inhibitor of mTOR, augmented thrombin-induced ICAM-1 expression. Inhibition of mTOR by this approach promoted whereas over-expression of mTOR inhibited thrombin-induced transcriptional activity of NF-kappa B, an essential regulator of ICAM-I transcription. Analysis of the NF-kappa B signaling pathway revealed that inhibition of mTOR potentiated I kappa B kinase activation resulting in a rapid and persistent phosphorylation of I kappa B alpha on Ser32 and Ser36, a requirement for I kappa B alpha degradation. Consistent with these data, we observed a more efficient and stable nuclear localization of RelA/p65 and, subsequently, the DNA binding activity of NF-kappa B by thrombin following mTOR inhibition. These data define a novel role of mTOR in down-regulating thrombin-induced ICAM-I expression in endothelial cells by controlling a delayed and transient activation of NF-kappa B.