Cytostatic effects of horseradish and thyroid peroxidase derived free radicals.

An otherwise noncytostatic flux of H2O2 from glucose and glucose oxidase became cytostatic to cultured Chinese Hamster Ovary (CHO) cells when horseradish or thyroid peroxidase was added to the culture medium. Electron spin resonance (ESR) measurements showed that one or more factors present in the culture medium promote the one-electron oxidation of a reduced nitroxide or glutathione in an H2O2/peroxidase-dependent process. Moreover, a reduced nitroxide conferred significant protection against the cytostatic effect of H2O2/peroxidase. Cytostatic effects were not only seen in the presence of the active H2O2/peroxidase system, but also in media which had been preexposed to H2O2/peroxidase but no longer contained an active H2O2 generating system. It is suggested that peroxidases oxidize one or more factors in tissue culture media to free radicals, which react with nearby components of cells or form toxic products, causing growth inhibition. If similar free radical precursors are present in tissue fluids, some of the toxicity of H2O2 in vivo may be due to peroxidase-mediated endogenous free radical generation.