Mechanisms of Anthracycline-Enhanced Reactive Oxygen Metabolism in Tumor Cells.

James H Doroshow

doi:10.1155/2019/9474823

Abstract

In this investigation, we examined the effect of anthracycline antibiotics on oxygen radical metabolism in Ehrlich tumor cells. In tumor microsomes and nuclei, doxorubicin increased superoxide anion production in a dose-dependent fashion that appeared to follow saturation kinetics; the apparent Km and Vmax for superoxide formation by these organelles was 124.9 μM and 22.6 nmol/min/mg, and 103.4 μM and 4.8 nmol/min/mg, respectively. In both tumor microsomes and nuclei, superoxide formation required NADPH as a cofactor, was accompanied by the formation of hydrogen peroxide, and resulted from the transfer of electrons from NADPH to the doxorubicin quinone by NADPH:cytochrome P-450 reductase (NADPH:ferricytochrome oxidoreductase, EC 1.6.2.4). Anthracycline antibiotics also significantly enhanced superoxide anion production by tumor mitochondria with an apparent Km and Vmax for doxorubicin of 123.2 μM and 14.7 nmol/min/mg. However, drug-stimulated superoxide production by mitochondria required NADH and was increased by rotenone, suggesting that the proximal portion of the electron transport chain in tumor cells was responsible for reduction of the doxorubicin quinone at this site. The net rate of drug-related oxygen radical production was also determined for intact Ehrlich tumor cells; in this system, treatment with doxorubicin produced a dose-related increase in cyanide-resistant respiration that was enhanced by changes in intracellular reducing equivalents. Finally, we found that in the presence of iron, treatment with doxorubicin significantly increased the production of formaldehyde from dimethyl sulfoxide, an indication that the hydroxyl radical could be produced by intact tumor cells following anthracycline exposure. These experiments suggest that the anthracycline antibiotics are capable of significantly enhancing oxygen radical metabolism in Ehrlich tumor cells at multiple intracellular sites by reactions that could contribute to the cytotoxicity of this class of drugs.

Highlights

The anthracycline antibiotics, including doxorubicin and daunorubicin, play an important role in the treatment of human leukemias and lymphomas as well as carcinomas of the breast [1]
To examine the mechanism of formaldehyde production and to verify that the evolution of formaldehyde from dimethyl sulfoxide (DMSO) was a measurement of ⋅OH formation, we investigated the effect of various oxygen radical scavengers on the level of formaldehyde produced by treatment of Ehrlich cells with doxorubicin
We have provided a comprehensive examination of the sites and mechanisms of anthracyclinestimulated oxy-radical production by Ehrlich carcinoma cells

Summary

Introduction

The anthracycline antibiotics, including doxorubicin and daunorubicin, play an important role in the treatment of human leukemias and lymphomas as well as carcinomas of the breast [1]. The major long-term toxicity of anthracycline therapy is a form of cumulative cardiac toxicity that may produce long-lived morbidity, especially in pediatric cancer patients receiving this class of drugs [2]. Redox cycling of the anthracycline quinone moiety by complex I of the cardiac electron transport chain [5] can damage intracellular lipid membranes in mitochondria as well as sodium and calcium transporters in the heart [6]. In addition to redox cycling of the quinone functionality of the anthracyclines, doxorubicin may form a potent drug-iron complex that enhances the formation of powerful oxidants with the chemical characteristics of the hydroxyl radical [7, 8].

Methods

Results

Conclusion