IKKalpha and IKKbeta function in TNFalpha-stimulated adhesion molecule expression in human aortic smooth muscle cells.
The role of NFkappaB and it's upstream kinases in regulating adhesion molecule expression in the smooth muscle of the vasculature remains controversial. We therefore examined the effect of blocking the NFkappaB pathway on TNFalpha-stimulated ICAM-1 and VCAM-1 expression in primary cultures of human aortic smooth muscle cells using an adenoviral wild-type IkappaB alpha construct (Ad.IkappaB alpha) and dominant-negative IKKalpha (Ad.IKKalpha+/-) and IKKbeta (Ad.IKKbeta+/-) constructs. Ad.IkappaB alpha treatment was found to block NFkappaB DNA-binding, and thereby completely prevent TNFalpha-stimulated ICAM-1 and VCAM-1 expression without influencing IKK activity. Ad.IKKbeta+/- treatment completely inhibited TNFalpha-stimulated IKK kinase activity, IkappaB alpha degradation and NFkappaB DNA-binding in addition to completely blocking TNFalpha-stimulated ICAM-1 and VCAM-1 expression. Ad.IKKalpha+/- treatment however had no detectable effect on NFkappaB DNA-binding or ICAM-1 and VCAM-1 expression. Our results demonstrate that TNFalpha-stimulated ICAM-1 and VCAM-1 expression in human aortic smooth muscle cells is NFkappaB-dependent, that IKKbeta is a suitable target for drug therapy and Ad.IKKbeta+/- an effective inhibitor of TNFalpha-stimulated ICAM-1 and VCAM-1 expression.
MacKenzie
Christopher J
CJ
Department of Physiology and Pharmacology, University of Strathclyde, John Arbuthnott Building, Strathclyde Institute for Biomedical Sciences, 27 Taylor Street, Glasgow, G4 0NR, Scotland, UK. christopher.mackenzie@strath.ac.uk
Ritchie
Elwyn
E
Paul
Andrew
A
Plevin
Robin
R
D001011:Aorta / Q000166:cytology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000201:enzymology; D004730:Endothelium, Vascular / Q000166:cytology; D004789:Enzyme Activation; D006801:Humans; D051550:I-kappa B Kinase / Q000502:physiology*; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis*; D032389:Myocytes, Smooth Muscle / Q000201:enzymology*; D016328:NF-kappa B / Q000378:metabolism; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis*
D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human