Effect of cobalt and chromium ions on bcl-2, bax, caspase-3, and caspase-8 expression in human U937 macrophages

The bcl-2 and caspase families of proteins play a central role in the modulation of apoptosis. The purpose of this study was to analyze the effect of Co2+ and Cr3+ ions on the expression of bcl-2, bax, caspase-3 and caspase-8 to better understand the mechanisms leading to ion-induced apoptosis in macrophages. U937 human macrophages were exposed to Co2+ and Cr3+ ions. The expression of proteins was measured by Western blot while caspase activities were measured by colorimetric assay. Results show that Co2+ ions inhibited bcl-2 expression with significant effect (p < 0.05) after 16 h and a maximal 52% inhibitory effect after 24 h. Co2+ stimulated bax expression with a significant stimulation (p < 0.05) after 8 h and a maximal 1.75-fold increase after 16 It. Co-2+ also stimulated the expression of the active fragment of caspase-3 as well as caspase-3 activity maximal increase after 24 It. Co2+ ions had no effect on caspase-8 expression or activity. Cr3+ ions inhibited bcl-2 expression with significant effect (p < 0.05) after 16h and a maximal 43% inhibitory effect after 24h. Cr3+ stimulated bax expression with significant stimulation (p < 0.01) after 8h and a maximal 2.25-fold increase after 24h. Cr3+ ions also stimulated the expression of the active fragments of caspase-3 and -8, as well as the activities of both proteases. The effect of Cr3+ ions on the expression of both caspase active fragments was maximal after 16h incubation. In conclusion, our results suggest that the modulation of the expression of proteins from the bcl-2 and the caspase families of proteins are implicated in the induction of macrophage apoptosis by Co2+ and Cr3+ ions. (C) 2003 Elsevier Ltd. All rights reserved.