Differential protective effects of varying degrees of hypoxia on the cytotoxicities of etoposide and bleomycin

Summary Oxygen is thought to be involved both directly and indirectly in the mechanisms of action of several anticancer agents. We studied the effects of various oxygen concentrations on the cytotoxicities of the following drugs: bleomycin (BLM), etoposide (VP-16), doxorubicin (DOX), and mitomycin C (MMC). Human sarcoma cells, MESSA, were exposed to drug for 1 h at one of several oxygen concentrations: less than 1%, 2.5%, 5%, 21%, and 95%. Cytotoxicity was assessed by cellular incorporation of ³H-thymidine into DNA 5 days after drug exposure. Control experiments varying oxygen concentration without drugs demonstrated toxicity only at the highest concentration (95%). Three different responses of drug sensitivity to varying oxygen tensions were observed. BLM, which has been shown to utilize oxygen as a substrate in generating free radicals and producing DNA scission, demonstrated a progressive increase in cytotoxicity over the entire range of increasing oxygen concentrations. This is consistent with the model of a BLM-cation-oxygen complex and catalytic reduction of oxygen. VP-16, which also produces DNA strand breakage but by interaction with topoisomerase II, exhibited a threshold response. VP-16 toxicity was ameliorated by anoxic conditions (less than 1% O₂), but not by oxygen concentrations of 2.5%–95%. The reason for this protective effect of anoxia with VP-16 is not clear. In contrast, acute anoxia had no effect on the cytotoxicities of DOX and MMC. We conclude that acute hypoxia protects cells from both BLM and VP-16 but that the nature of that protection is different. VP-16 toxicitiy is blunted only by severe anoxia, wheaeas BLM exhibits a dose response effect over the entire range of oxygen concentrations.Supported by NIH grant CA-27478 from the US Department of Health and Human Services, and by the American Lung Association. Dr. Sikic is a recipient of a Faculty Development Award in Clinical Pharmacology from the Pharmaceutical Manufacturer's Association Foundation.