Redox cycling of bleomycin-Fe(III) by an NADH-dependent enzyme, and DNA damage in isolated rat liver nuclei.

Isolated rat liver nuclei were incubated aerobically with bleomycin (BLM) and FeCl3 in the presence of NADH. An increase in NADH and oxygen consumption was observed accompanied by DNA cleavage as shown by gel electrophoresis. Malondialdehyde (MDA) was also formed, which partly derived from DNA indicating an oxidative cleavage mechanism. BLM and NADH were obligatory to provide these effects, whereas FeCl3 could be omitted, without a complete loss of the activities mentioned above. This was explained by the presence of some iron in the nuclei. NADPH was consumed to a lesser extent compared to NADH and was less effective with respect to O2 consumption and MDA formation. It could be excluded that mitochondrial or microsomal contaminations in nuclear preparations were responsible for the effects observed. The results suggest that the BLM-Fe(III)-complex can be repeatedly reduced (redox cycled) by NADH- (and NADPH-) dependent reductases of liver nuclei to BLM-Fe(II) which is known to form reactive oxygen species and to damage DNA. It is concluded that the enzymatic reduction of a BLM-metal complex in the cell nucleus may be an essential step in the cytotoxic activity of bleomycin.