The role of protein kinase C in the induction of VCAM‐1 expression on human umbilical vein endothelial cells

Deisher, Theresa A.; Haddix, Terri; Montgomery, Kevin F.; Pohlman, Timothy H.; Kaushansky, Kenneth; Harlan, John M.

doi:10.1016/0014-5793(93)80354-w

Cited by 61 publications

(40 citation statements)

References 20 publications

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“…Furthermore, oleate also inhibited VCAM-1 expression induced by PMA, an agent able to induce endothelial activation that bypasses cell surface receptors and likely acting through direct activation of protein kinase C, possibly even bypassing NF-B activation. 34 We also confirm, in line with what we reported for other more unsaturated fatty acids, 17 but contrary to what has been reported by others for DHA, 19 that the effect of oleate is not restricted to VCAM-1 expression but also pertains to other inducible adhesion molecules, suggesting a mechanism on a common pathogenetic transducer of endothelial activation. Together with the data on the inhibition of VCAM-1 mRNA accumulation obtained herein by Northern blot analysis and the notion that the regulation of adhesion molecule expression appears to be to largely transcriptional, 20 we postulate transcriptional interference by oleate, possibly mediated by inhibition of the activation of the transcription factor NF-B.…”

Section: Discussionsupporting

confidence: 91%

Oleic Acid Inhibits Endothelial Activation

Carluccio¹,

Massaro²,

Bonfrate³

et al. 1999

ATVB

209

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Abstract-Because oleic acid is implicated in the antiatherogenic effects attributed to the Mediterranean diet, we investigated whether this fatty acid can modulate endothelial activation, ie, the concerted expression of gene products involved in leukocyte recruitment and early atherogenesis. We incubated sodium oleate with human umbilical vein endothelial cells for 0 to 72 hours, followed by coincubation of oleate with human recombinant tumor necrosis factor, interleukin (IL)-1␣, IL-1␤, IL-4, Escherichia coli lipopolysaccharide (LPS), or phorbol 12-myristate 13-acetate for a further 6 to 24 hours. The endothelial expression of vascular cell adhesion molecule-1 (VCAM-1), E-selectin, and intercellular adhesion molecule-1 was monitored by cell surface enzyme immunoassays or flow cytometry, and steady-state levels of VCAM-1 mRNA were assessed by Northern blot analysis. At 10 to 100 mol/L for Ͼ24 hours, oleate inhibited the expression of all adhesion molecules tested. After a 72-hour incubation with oleate and a further 16-hour incubation with oleate plus 1 g/mL LPS, VCAM-1 expression was reduced by Ͼ40% compared with control. Adhesion of monocytoid U937 cells to LPS-treated endothelial cells was reduced concomitantly. Oleate also produced a quantitatively similar reduction of VCAM-1 mRNA levels on Northern blot analysis and inhibited nuclear factor-B activation on electrophoretic mobility shift assays. Incubation of endothelial cells with oleate for 72 hours decreased the relative proportions of saturated (palmitic and stearic) acids in total cell lipids and increased the proportions of oleate in total cell lipids without significantly changing the relative proportions of polyunsaturated fatty acids. Although less potent than polyunsaturated fatty acids in inhibiting endothelial activation, oleic acid may contribute to the prevention of atherogenesis through selective displacement of saturated fatty acids in cell membrane phospholipids and a consequent modulation of gene expression for molecules involved in monocyte recruitment. (Arterioscler Thromb Vasc Biol. 1999;19:220-228.)

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Section: Discussionsupporting

confidence: 91%

Oleic Acid Inhibits Endothelial Activation

Carluccio¹,

Massaro²,

Bonfrate³

et al. 1999

ATVB

209

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“…The various PKC isoforms have been shown to differ in their spatial distribution both in resting endothelial cells and in response to extracellular mediators (12). PKCs have been implicated in multiple endothelial cell functions, including expression of adhesion molecules (14), and endothelin-1 (15), proliferative response to growth factors (16), shear stress signaling (17), and angiogenesis (18).…”

Section: Vascular Adhesion Molecule-1 (Vcam-1)mentioning

confidence: 99%

Thrombin Stimulation of Vascular Adhesion Molecule-1 in Endothelial Cells Is Mediated by Protein Kinase C (PKC)-δ-NF-κB and PKC-ζ-GATA Signaling Pathways

Minami

Abid

Zhang

et al. 2003

Journal of Biological Chemistry

106

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We recently demonstrated that thrombin induces the expression of vascular adhesion molecule-1 (VCAM-1) in endothelial cells by an NF-B-and GATA-dependent mechanism. In the present study, we describe the signaling pathways that mediate this response. Thrombin stimulation of the VCAM-1 gene and promoter in human umbilical vein endothelial cells was inhibited by preincubation with the phosphatidylinositol 3-kinase inhibitor, LY294002, the protein kinase C (PKC)-␦ inhibitor, rottlerin, a PKC-peptide inhibitor, or by overexpression of dominant negative (DN)-PKC-. In electrophoretic mobility shift assays, thrombin-mediated induction of NF-B p65 binding to two NF-B motifs in the upstream promoter region of VCAM-1 was blocked by LY294002 and rottlerin, whereas the inducible binding of GATA-2 to a tandem GATA motif was inhibited by LY294002 and the PKC-peptide inhibitor. In co-transfection assays, thrombin stimulation of a minimal promoter containing multimerized VCAM-1 NF-B sites was inhibited by DN-PKC-␦ but not DN-PKC-. In contrast, thrombin-mediated transactivation of a minimal promoter containing tandem VCAM-1 GATA motifs was inhibited by DN-PKC-but not DN-PKC-␦. Finally, thrombin failed to induce VCAM-1 expression in vascular smooth muscle cells. Taken together, these data suggest that the endothelial cell-specific effect of thrombin on VCAM-1 expression involves the coordinate activity of PKC-␦-NF-B and PKC--GATA signaling pathways. Vascular adhesion molecule-1 (VCAM-1)1 is a 110-kDa cell surface glycoprotein that is expressed in cytokine-activated endothelial cells (1). The VCAM-1 promoter, originally cloned and characterized in cultured endothelial cells (2), represents a potentially valuable tool for dissecting the molecular mechanisms of endothelial cell activation. Several studies have demonstrated the importance of two tandem NF-B elements located at position Ϫ77 and Ϫ63, relative to the transcriptional start site, in transducing the response to inflammatory mediators (2-4). Other studies have provided evidence for the role of co-stimulators in mediating cytokine response, including Sp1 (5), activating protein-1 (6), and interferon regulatory factor-1 (7).In a recent report, we showed that the incubation of endothelial cells with thrombin or the PAR-1 agonist, thrombin receptor activation peptide (TRAP), resulted in increased VCAM-1 mRNA levels and VCAM-1 promoter activity (8). Not surprisingly, a mutation of the tandem NF-B motif blocked the response to thrombin. Moreover, in electrophoretic mobility shift assays, thrombin induced the binding of p65 homodimers to the two adjacent NF-B sites in the VCAM-1 promoter. A more interesting finding was that a mutation of a tandem GATA motif (located at Ϫ244 and Ϫ259) also attenuated the thrombin response. Consistent with these results, thrombin treatment resulted in increased binding of GATA-2 to the VCAM-1 promoter. These data suggested that thrombin-mediated induction of VCAM-1 involves the coordinate activity of NF-B p65 and GATA-2 and raised new questions as to...

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“…Furthermore, treatment of HUVEC for 48 h with PMA, which downregulates PKC (Ritchie et al, 1991;Deisher et al, 1993a), did not prevent re-induction of E-selectin by TNFa, nor did it affect the potency of genistein to inhibit this re-induced expression. Taken together these results confirm that cytokineinduced E-selectin expression is PKC-independent (Ritchie et al, 1991) and show that inhibition of expression by genistein is not via PKC.…”

Section: U T T T T T U T T T T Tmentioning

confidence: 99%

Effects of protein tyrosine kinase inhibitors on cytokine‐induced adhesion molecule expression by human umbilical vein endothelial cells

May

Wheeler‐Jones

Pearson

1996

British J Pharmacology

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. Endothelial cells can be stimulated by the pro‐inflammatory cytokines interleukin (IL)‐1α and tumour necrosis factor (TNF)α to express the leukocyte adhesion molecules E‐selectin, vascular cell adhesion molecule (VCAM)‐1 and intercellular adhesion molecule (ICAM)‐1 but the intracellular signalling mechanisms leading to this expression are incompletely understood. We have investigated the role of protein tyrosine kinases (PTK) in adhesion molecule expression by cytokine‐activated human umbilical vein endothelial cells (HUVEC) using the PTK inhibitors genistein and herbimycin A, and the protein tyrosine phosphatase (PTP) inhibitor sodium orthovanadate. . Maximal E‐selectin expression induced by incubation of HUVEC for 4 h with IL‐1α (100 u ml−1) and TNFα (100 u ml−1) was dose‐dependently inhibited by genistein and herbimycin A. Although similar effects were seen on phorbol 12‐myristate, 13‐acetate (PMA)‐induced expression, this was not due to inhibition of protein kinase C (PKC) activity as the selective inhibitors of PKC, bisindolylmaleimide (BIM), Ro31‐7549 or Ro31‐8220 did not affect IL‐1α‐ or TNFα‐induced E‐selectin expression at concentrations which maximally inhibited PMA‐induced expression. . Genistein inhibited VCAM‐1 expression induced by incubation of HUVEC for 24 h with TNFα or IL‐1α whereas it did not affect ICAM‐1 expression induced by 24 h incubation with either of these cytokines. Herbimycin A inhibited both VCAM‐1 and ICAM‐1 expression induced by TNFα. . Basal expression of E‐selectin, VCAM‐1 and ICAM‐1 was dose‐dependently enhanced by sodium orthovanadate. In contrast, vanadate differentially affected TNFα‐induced expression of these molecules with maximal E‐selectin and ICAM‐1 expression being slightly enhanced and VCAM‐1 expression dose‐dependently reduced. . We also studied the effects of PTK and PTP inhibitors on adhesion of the human pre‐myeloid cell line U937 to TNFα‐stimulated HUVEC. Adhesion of U937 cells to HUVEC pretreated for 4 or 24 h with TNFα was dose‐dependently inhibited by genistein and herbimycin A but unaffected by daidzein. Adhesion of U937 cells after 4 h was partially inhibited by blocking antibodies against both E‐selectin and VCAM‐1 but after 24 h was only inhibited by anti‐VCAM‐1. . Sodium orthovanadate had no effect on TNFα‐induced U937 adhesion but dose‐dependently enhanced adhesion to unstimulated HUVEC. Vanadate‐induced adhesion was inhibited by an antibody against VCAM‐1. . These results demonstrate that PTK‐mediated phosphorylation events are important for the regulation of adhesion molecule expression by human endothelial cells, and additionally show that PTK inhibitors differentially affect upregulation of different adhesion molecules, implicating divergent regulatory pathways for cytokine‐induced adhesion molecule expression.

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The role of protein kinase C in the induction of VCAM‐1 expression on human umbilical vein endothelial cells

Cited by 61 publications

References 20 publications

Oleic Acid Inhibits Endothelial Activation

Oleic Acid Inhibits Endothelial Activation

Thrombin Stimulation of Vascular Adhesion Molecule-1 in Endothelial Cells Is Mediated by Protein Kinase C (PKC)-δ-NF-κB and PKC-ζ-GATA Signaling Pathways

Effects of protein tyrosine kinase inhibitors on cytokine‐induced adhesion molecule expression by human umbilical vein endothelial cells

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