Proteasome inhibitors block VCAM-1 and ICAM-1 gene expression in endothelial cells without affecting nuclear translocation of nuclear factor-kappa B

Eur J Immunol. 1996 Apr;26(4):839-45. doi: 10.1002/eji.1830260417.

Abstract

Endothelial cells play a major role in recruiting leukocytes to sites of inflammation. This is accomplished, at least in part, by up-regulation of cell surface adhesion molecules, including VCAM-1 and ICAM-1, in response to cytokines. In this report, we investigated the role of the proteasome complex in mediating the interleukin (IL)- 1 beta induction of VCAM-1 and ICAM-1 gene expression in human endothelial cells. We present evidence that a proteasome inhibitor, n-acetyl-leucinyl-leucinyl-norleucinal (norLEU), as well as specific protease inhibitors, n-tosyl-Lys-chloromethylketone and N-tosyl-Phe-chloromethylketone, blocked IL-1 beta induction of VCAM-1 and ICAM-1 promoter-driven reporter gene expression in stably transfected endothelial cells. These inhibitors also blocked cytokine induced cell surface expression of VCAM-1 and ICAM-1 by human umbilical vein endothelial cells. As expected, the protease inhibitors blocked the activation of nuclear factor (NF)-kappa B in response to IL-1 beta stimulation. In contrast, norLEU did not prevent IL-1 beta-induced nuclear translocation of NF-kappa B. The effects of norLEU were specific because it did not inhibit the IL-1 beta induction of plasminogen activator inhibitor type 1 gene expression. This study demonstrates that inhibition of the proteolytic activity of the proteasome blocks IL-1 beta induction of VCAM-1 and ICAM-1 gene expression in human endothelial cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Biological Transport / drug effects
  • Calpain / antagonists & inhibitors
  • Calpain / physiology*
  • Cell Nucleus / metabolism*
  • Cells, Cultured
  • Cysteine Endopeptidases / physiology*
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism*
  • Gene Expression Regulation / drug effects*
  • Humans
  • Intercellular Adhesion Molecule-1 / biosynthesis*
  • Intercellular Adhesion Molecule-1 / genetics
  • Interleukin-1 / pharmacology
  • Leupeptins / pharmacology*
  • Molecular Sequence Data
  • Multienzyme Complexes / physiology*
  • NF-kappa B / metabolism*
  • Plasminogen Activator Inhibitor 1 / biosynthesis
  • Plasminogen Activator Inhibitor 1 / genetics
  • Protease Inhibitors / pharmacology
  • Proteasome Endopeptidase Complex
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Recombinant Fusion Proteins / metabolism
  • Tosyllysine Chloromethyl Ketone / pharmacology*
  • Tosylphenylalanyl Chloromethyl Ketone / pharmacology*
  • Transfection
  • Vascular Cell Adhesion Molecule-1 / biosynthesis*
  • Vascular Cell Adhesion Molecule-1 / genetics

Substances

  • Interleukin-1
  • Leupeptins
  • Multienzyme Complexes
  • NF-kappa B
  • Plasminogen Activator Inhibitor 1
  • Protease Inhibitors
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Vascular Cell Adhesion Molecule-1
  • acetyl-leucinyl-leucinyl-methional
  • acetylleucyl-leucyl-norleucinal
  • Intercellular Adhesion Molecule-1
  • Tosyllysine Chloromethyl Ketone
  • Tosylphenylalanyl Chloromethyl Ketone
  • Calpain
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex