DNA Cross-Linking in Mammalian Cells by Pyrrolizidine Alkaloids: Structure-Activity Relationships
Toxicology and Applied Pharmacology
Pyrrolizidine alkaloids (PAs) are common constituents of many species of flowering plants which posses carcinogenic as well as anticarcinogenic activityin vivo. Pyrrolizidine alkaloids are genotoxic in various short-term assays. The mechanisms by which these compounds exert these effects is still unclear. In this study, we characterized the ability of eight bifunctional PAs, with differing stereochemistry and functional groups, to cross-link cellular DNA in cultured bovine kidney epithelial cells. PAs representative of three major structural classes, the macrocycles (seneciphylline, riddelline, retrorsine, senecionine, monocrotaline), the open diesters (heliosupine, latifoline), and pyrrolizidine base (retronecine) were cultured for 2 hr with cells and an external metabolizing system. Every PA induced DNA cross-links which consisted primarily of proteinase-sensitive cross-links (DPC), but also to a smaller extent, DNA interstrand cross-links (ISC). None of the PAs induced detectable amounts of DNA single-strand breaks. The PAs which produced DPC and/or ISC (ranked from most potent to least) were: seneciphylline (DPC > ISC); riddelline (DPC > ISC); retrorsine (DPC > ISC); senecionine (DPC > ISC); heliosupine (DPC > ISC); monocrotaline (ISC = DPC); latifoline (DPC > ISC); and retronecine (ISC > DPC). Although the PAs induced DNA cross-linking to varying degrees, cell viabilities for all treatment groups were >90% as determined by trypan blue dye exclusion. Since the cross-linking ability of these PAs paralleled their ability to inhibit colony formation, cross-link formation may be involved in the biological activity of these compounds. Two structural determinants of biological activity appear to be the presence of both a macrocyclic necic acid ester and an ά,β-unsaturated ester function since the cross-linking ability of seneciphylline, riddelline, retrorsine, and senecionine far exceeded that of monocrotaline, heliosupine, latifoline, and retronecine. In addition, the stereochemical orientation of the ester linkage was found to have no effect on bm logicaI activia.
Hincks, J.R., H.Y. Kim, H.J. Segall, R.J. Molyneaux, F.R. Stermitz and R.A. Coulombe, Jr. (1991). DNA cross-linking in mammalian cells by pyrrolizidine alkaloids: structure-activity relationships Toxicol. Appl. Pharmacol. 111:90-98.