Apoptosis or programmed cell death was induced in the human promyelocytic leukemia cell line HL-60 by UV irradiation or treatment with cytotoxic drugs (etoposide, camptothecin, melphalan or chlorambucil). These treatments caused a rapid increase in intracellular peroxide levels. Preincubation of HL-60 cells with the hydrogen peroxide-scavenging enzyme catalase (500 U/ml) inhibited apoptosis due to UV irradiation or low concentrations of camptothecin, etoposide or melphalan, but did not protect against higher concentrations. In contrast, superoxide anion levels in the cells remained unchanged upon treatment with cytotoxic drugs, while UV irradiation led to a transient doubling in superoxide levels. Exogenous superoxide dismutase (400 U/ml) provided modest protection against UV irradiation and had no effect on cytotoxic drug-induced apoptosis. The results suggest that both hydrogen peroxide and superoxide anion may be involved in the induction of apoptosis by UV irradiation. On the other hand, while exposure to cytotoxic drugs induces a large increase in intracellular peroxide levels, catalase is able to protect the cells from apoptosis only when low concentrations of these compounds are used, thus indicating the involvement of other factors in this process, particularly at higher drug concentrations.