COPPER-INDUCED TISSUE FACTOR EXPRESSION IN HUMAN MONOCYTIC THP-1 CELLS AND ITS INHIBITION BY ANTIOXIDANTS

Background Transition metals such as copper are known to initiate Gee radical formation and lipid peroxidation. Recent reports suggest that intracellular reactive oxygen intermediates can induce the transcription of a number of important genes. The present study examines the effects of copper and iron on the ability of monocytic cells to synthesize and express tissue factor, the potent procoagulant factor. Methods and Results Exposure of human monocytic THP-1 cells to 5 to 10 mu mol/L Cu2+ led to cell damage and the expression of tissue factor activity to levels up to 70 times higher than control, as measured by a single-stage plasma coagulation assay. These effects were seen only in the presence of a lipophilic chelating agent, 8-hydroxyquinoline, suggesting that intracellular transport of Cu2+ was required. The effects of Cu2+ were mimicked by ceruloplasmin but not by Fe3+ or hemin. The induction of tissue factor activity by Cu2+ was slow in onset (6 hours) but sustained (24 hours) and was accompanied by increased tissue factor mRNA levels, measured by reverse transcription/polymerase chain reaction after annealing with oligomer primers. Increases in tissue factor protein, measured by a specific immunoassay, also occurred but were smaller than those in activity. Cu2+, therefore, appears to act at both the transcriptional and posttranslational levels. The effects of Cu2+ were inhibited by a number of lipophilic antioxidants, including probucol, vitamin E, butylated hydroxytoluene, and a 21-aminosteroid, U74389G. Conclusions Exposure of monocytes to oxidizing conditions may lead to the expression of high levels of tissue factor activity, with accompanying risk for disseminated intravascular coagulation, and this may be inhibited by lipophilic antioxidants.