DNA Adduct Formation by the Anticancer Drug Ellipticine and Its Hydroxy Derivatives in Human Breast Adenocarcinoma MCF-7 Cells

Abstract

The cytotoxicity of the antineoplastic agent ellipticine and its 9- and 7-hydroxylated metabolites to human breast adenocarcinoma MCF-7 cells and their ability to generate DNA adducts in these cancer cells were investigated. Ellipticine and its 9-hydroxylated metabolite were found to be toxic to MCF-7 cells with IC50values of 1.25 and 3.25 μmol l-1for ellipticine and 9-hydroxyellipticine, respectively. In contrast, no toxicity to these cancer cells was detectable for 7-hydroxyellipticine. The nuclease P1 version of the32P-postlabeling assay yielded a pattern of ellipticine-DNA adducts with two major and one minor adducts in MCF-7 cells, similar to the pattern of adducts detected in DNA reacted with ellipticine and the reconstituted cytochrome P450 enzyme systemin vitroand in DNAin vivo. The identity of two major adducts formed in DNA of MCF-7 cells with those formed by cytochrome P450-mediated ellipticine activationin vitrowas confirmed by HPLC of the isolated adducts. 9-Hydroxyellipticine was also capable of inducing DNA adducts in MCF-7 cells, but to a lesser extent. In addition, the adducts generated by 9-hydroxyellipticine were different from those generated by ellipticine. Negligible levels of DNA adducts were detectable in DNA of MCF-7 cells exposed to 7-hydroxyellipticine. The results presented here are the first report showing the formation of covalent DNA adducts with ellipticine in human breast cancer cells in culture, and suggest the formation of covalent DNA adducts as a new mode of antitumor action of ellipticine in breast cancer.