Inhibitors of topoisomerase II prevent cytokine-induced expression of vascular cell adhesion molecule-1, while augmenting the expression of endothelial leukocyte adhesion molecule-1 on human umbilical vein endothelial cells.
Cytokine stimulation of human umbilical vein endothelial cells (HUVE) induces surface expression of the adhesion molecules vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and endothelial leukocyte adhesion molecule-1 (E-selectin). We previously found that induction of adhesion molecule expression in HUVE is regulated, at least in part, by protein kinase C (PKC) activation, although this is not associated with the expected translocation of PKC from the cytosolic to the particulate fraction. We therefore investigated potential nuclear targets for PKC. Topoisomerase II is localized to the nuclear matrix and has been shown to be phosphorylated, both in vitro and in vivo, by PKC. In HUVE, the topoisomerase II selective inhibitors novobiocin, nalidixic acid, and etoposide prevented cytokine-induced VCAM-1 surface expression, but not E-selectin or ICAM-1 surface expression. Similarly, novobiocin and nalidixic acid reduced the accumulation of VCAM-1 mRNA in response to tumor necrosis factor-alpha treatment of HUVE. The inhibitory effect of the topoisomerase II inhibitors on VCAM-1 expression was not due to non-specific toxicity, as protein synthesis, measured by trichloroacetic acid precipitation of 35S-methionine labeled proteins, and transcription, determined by beta-actin mRNA levels, were not decreased. In contrast to the observed reduction of VCAM-1 mRNA accumulation and surface protein expression, inhibition of topoisomerase II activity enhanced E-selectin mRNA accumulation and surface protein expression in response to tumor necrosis factor-alpha stimulation of HUVE. This work demonstrates that topoisomerase II activity may differentially regulate the expression of adhesion molecules on HUVE.
Deisher
T A
TA
Department of Medicine, University of Washington Medical Center, Seattle 98195.
Kaushansky
K
K
Harlan
J M
JM
D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000235:genetics*; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D005047:Etoposide / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D009268:Nalidixic Acid / Q000494:pharmacology; D009675:Novobiocin / Q000494:pharmacology; D014176:Protein Biosynthesis; D011493:Protein Kinase C / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D059005:Topoisomerase II Inhibitors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D012333:RNA, Messenger; D059005:Topoisomerase II Inhibitors; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D009675:Novobiocin; D009268:Nalidixic Acid; D005047:Etoposide; D004247:DNA; D011493:Protein Kinase C