TRANSCRIPTION FACTOR DECOY FOR NUCLEAR FACTOR-KAPPA-B INHIBITS TUMOR NECROSIS FACTOR-ALPHA-INDUCED EXPRESSION OF INTERLEUKIN-6 AND INTRACELLULAR ADHESION MOLECULE-1 IN ENDOTHELIAL-CELLS

Background Several cytokines and adhesion molecules released from endo thelium play an important role in inflammation, immune responses, and probably atherogenesis. Objective To determine whether the transcripti on factor nuclear factor-kappa B mediated expression of these genes in volved in the inflammatory response of endothelial cells to tumor necr osis factor-alpha, by using transcription factor decoy oligodeoxynucle otides. Design and methods We first transfected fluorescein isothiocya nate (FITC)-labeled double-stranded oligodeoxynucleotides into endothe lial cells by a cationic liposome-mediated method of gene transfer. We then confirmed that the decoy oligodeoxynucleotides could block bindi ng of nuclear factor-kappa B to its specific cis element effectively. In addition, we transfected the reporter gene chloramphenicol acetyltr ansferase driven by three repeated nuclear factor-kappa B binding sequ ences in the promoter and enhancer region. Results FITC-labeled oligod eoxynucleotides were detected in the nuclei of approximately 70% of th e total cells. Tumor necrosis factor-stimulated expression of chloramp henicol acetyltransferase was partially inhibited by transfection of n uclear factor-kappa B decoy oligodeoxynucleotides, but not by transfec tion of scrambled oligodeoxynucleotides. Also nuclear factor-kappa B d ecoy oligodeoxynucleotides but not scrambled oligodeoxynucleotides inh ibited tumor necrosis factor-induced expression of interleukin-6 and i ntracellular adhesion molecule-1 both at the messenger RNA and at prot ein level (assessed by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay). Conclusion Out results demons trate that nuclear factor-kappa B decoy oligodeoxynucleotides transfec ted by cationic liposome method inhibited tumor necrosis factor-induce d expression of interleukin-6 and intracellular adhesion molecule-1 in endothelial cells, I Hypertens 16:993-1000 (C) 1998 Lippincott-Raven Publishers.