The mechanism of action of the clinically used iron(III) chelator, desferrioxamine (DFO), on preventing iron (Fe) uptake from transferrin (Tf) has been investigated using the human melanoma cell line SK-MEL-28. This investigation was initiated due to the paucity of information on the mechanisms of action of DFO in neoplastic cells and because recent studies have suggested that DFO may be a useful antitumor agent. The effect of DFO was dependent on incubation time. After a 2-h incubation, DFO acted like the extracellular chelators, EDTA and diethylenetriaminepentaacetic acid, because there was little inhibition of 59Fe uptake from Tf. In contrast, after a 24-h incubation, DFO (0.5 mm) efficiently reduced internalized 59Fe uptake from Tf to 18% of the control value. These observations suggested the existence of a kinetic block to the entry of the apochelator to intracellular Fe pools and/or to the exit of the DFO-59Fe complex. Indeed, cellular fractionation demonstrated that, in contrast to the decrease in the percentage of 59Fe in the ferritin and membrane fractions, a marked increase in the percentage of 59Fe present in the ferritin-free cytosol occurred. These observations suggested an accumulation of the DFO-59Fe complex within the cell. The highly lipophilic Fe chelator, pyridoxal isonicotinoyl hydrazone, was far more effective than DFO at preventing 59Fe uptake from Tf, illustrating the importance of membrane permeability for effective Fe chelation. Desferrioxamine at a concentration of 1 mm decreased internalized 125I-Tf uptake to 70% of the control. However, the decrease in 59Fe uptake observed could only be partially accounted for by a decrease in Tf uptake, and it appeared that DFO was chelating 59Fe at an intracellular site consistent with the transit Fe pool. The results are discussed in the context of the use of Fe chelators as effective antineoplastic agents.
Supported by Grant 880524 (to E. B.) and a Biomedical Postgraduate Scholarship (to D. R. R.) from the National Health and Medical Research Council of Australia.