Urokinase mediates endothelial cell survival via induction of the X-linked inhibitor of apoptosis protein

Blood. 2009 Feb 5;113(6):1383-90. doi: 10.1182/blood-2008-06-164210. Epub 2008 Oct 23.

Abstract

Urokinase-type plasminogen activator (uPA) additionally elicits a whole array of pro-angiogenic responses, such as differentiation, proliferation, and migration. In this study, we demonstrate that in endothelial cells uPA also protects against apoptosis by transcriptional up-regulation and partially by mRNA stabilization of inhibitor of apoptosis proteins, most prominently the X-linked inhibitor of apoptosis protein (XIAP). The antiapoptotic activity of uPA was dependent on its protease activity, the presence of uPA receptor (uPAR) and low-density lipoprotein receptor-related protein (LRP), but independent of the phosphatidylinositol 3 (PI3) kinase pathway, whereas vascular endothelial growth factor (VEGF)-induced antiapoptosis was PI3 kinase dependent. uPA-induced cell survival involved phosphorylation of p21-activated kinase 1 (Pak1) and the IkappaB kinase alpha that leads to nuclear factor kappaB (NF-kappaB) p52 activation. Indeed, blocking NF-kappaB activation by using specific NF-kappaB inhibitors abolished uPA-induced cell survival as it blocked uPA-induced XIAP up-regulation. Furthermore, down-regulating XIAP expression by small interfering RNA (siRNA) significantly reduced uPA-dependent endothelial cell survival. This mechanism is also important for VEGF-induced antiapoptosis because VEGF-dependent up-regulation of XIAP was found defective in uPA(-/-) endothelial cells. This led us to conclude that uPA is part of a novel NF-kappaB-dependent cell survival pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • Cell Survival
  • Cells, Cultured
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism*
  • Humans
  • I-kappa B Kinase / metabolism
  • Immunoblotting
  • Immunoenzyme Techniques
  • Immunoprecipitation
  • Low Density Lipoprotein Receptor-Related Protein-1 / metabolism
  • Mice
  • Mice, Knockout
  • NF-kappa B / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphorylation
  • RNA Stability
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / pharmacology
  • Receptors, Urokinase Plasminogen Activator / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Urokinase-Type Plasminogen Activator / physiology*
  • Vascular Endothelial Growth Factor A / metabolism
  • X-Linked Inhibitor of Apoptosis Protein / antagonists & inhibitors
  • X-Linked Inhibitor of Apoptosis Protein / genetics
  • X-Linked Inhibitor of Apoptosis Protein / metabolism*
  • p21-Activated Kinases / metabolism

Substances

  • Low Density Lipoprotein Receptor-Related Protein-1
  • NF-kappa B
  • RNA, Messenger
  • RNA, Small Interfering
  • Receptors, Urokinase Plasminogen Activator
  • Vascular Endothelial Growth Factor A
  • X-Linked Inhibitor of Apoptosis Protein
  • Phosphatidylinositol 3-Kinases
  • p21-Activated Kinases
  • I-kappa B Kinase
  • Urokinase-Type Plasminogen Activator