Involvement of reactive oxygen species in aclarubicin-induced differentiation and invasiveness of HL-60 leukemia cells.

Aclarubicin (ACLA), which belongs to the anthracycline class of antineoplastic agents, has been demonstrated as a differentiating agent of human leukemia, including HL-60 cells. We report here on the incidence of ACLA-induced differentiation on matrix metalloproteinase (MMP) expression and cell invasiveness. The aim of this study was to investigate the involvement of reactive oxygen species (ROS) as mediators of ACLA-induced effects. By using a fluorescent probe, we showed that subtoxic (i.e. differentiating) concentration of ACLA generate reactive oxygen species in HL-60 cells. ACLA-differentiated cells exhibited an increased proMMP-9 secretion which has been observed by gelatin zymography and immunoassay. Antioxidants were able to inhibit ACLA-induced differentiation and proMMP-9 secretion. Furthermore, RT-PCR showed that ACLA increased MMP-9 and tissue inhibitor of MMP (TIMP-1) expression in a ROS-dependent manner. In addition, the migration and invasion capacities of HL-60 cells were enhanced by ACLA treatment, but only partially reversed by antioxidants. Altogether, these results evidenced ROS as messengers of ACLA-induced differentiation and MMP-9 expression.