title: DNA cross-linking in mammalian cells by pyrrolizidine alkaloids: structure-activity relationships.
abstract: Pyrrolizidine alkaloids (PAs) are common constituents of many species of flowering plants which possess carcinogenic as well as anticarcinogenic activity in 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 greater than ISC); riddelline (DPC greater than ISC); retrorsine (DPC greater than ISC); senecionine (DPC greater than ISC); heliosupine (DPC greater than ISC); monocrotaline (ISC = DPC); latifoline (DPC greater than ISC); and retronecine (ISC greater than DPC). Although the PAs induced DNA cross-linking to varying degrees, cell viabilities for all treatment groups were greater than 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 alpha,beta-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 biological activity.
authors.affiliation: Department of Veterinary Science, Utah State University, Logan 84322-4620.
authors.forename: J R
authors.initials: JR
authors.lastname: Hincks
authors.forename: H Y
authors.initials: HY
authors.lastname: Kim
authors.forename: H J
authors.initials: HJ
authors.lastname: Segall
authors.forename: R J
authors.initials: RJ
authors.lastname: Molyneux
authors.forename: F R
authors.initials: FR
authors.lastname: Stermitz
authors.forename: R A
authors.initials: RA
authors.lastname: Coulombe
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects* / Q000378:metabolism; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D007668:Kidney / Q000166:cytology; D011763:Pyrrolizidine Alkaloids / Q000494:pharmacology*; D013234:Stem Cells / Q000187:drug effects
chemical_list: D003432:Cross-Linking Reagents; D011763:Pyrrolizidine Alkaloids; D004247:DNA