Radiosensitizing and cytocidal effects of misonidazole: evidence for separate modes of action

Hypoxic BP-8 murine sarcoma cells were exposed to misonidazole and/or radiation and the kinetics and extent of cell death were evaluated with the [125I]iododeoxyuridine-prelabeling assay. Cell death after treatment with lethal doses of misonidazole was rapid and essentially complete within 2 or 3 days after drug exposure. In contrast, radiation death became apparent only after a delay period of 4 days and was complete by Day 10 after irradiation. Radiosensitization by short exposures to sublethal doses of misonidazole affected only the delayed component of cell death, that is, the radiation component of death. In experiments involving sequential radiation and drug treatment, prior irradiation of cells did not enhance the direct cytocidal effects of misonidazole, as evidenced by the fact that the early component of cell death was equal in control and preirradiated cells. However, postirradiation treatment with misonidazole did enhance the delayed radiation component of cell death. These results suggest that radiosensitization and direct killing by misonidazole are two distinct phenomena mediated by different cellular mechanisms, and radiosensitization by misonidazole represents a two-component effect composed of true dose modification and dose additive damage interactions, but these additive effects must occur at a site different from the cellular structure responsible for direct drug-induced cell death.