title: Nucleolar targeting by platinum: p53-independent apoptosis follows rRNA inhibition, cell-cycle arrest, and DNA compaction.
abstract: TriplatinNC is a highly positively charged, substitution-inert derivative of the phase II clinical anticancer drug, BBR3464. Such substitution-inert complexes form a distinct subset of polynuclear platinum complexes (PPCs) interacting with DNA and other biomolecules through noncovalent interactions. Rapid cellular entry is facilitated via interaction with cell surface glycosoaminoglycans and is a mechanism unique to PPCs. Nanoscale secondary ion mass spectrometry (nanoSIMS) showed rapid distribution within cytoplasmic and nucleolar compartments, but not the nucleus. In this article, the downstream effects of nucleolar localization are described. In human colon carcinoma cells, HCT116, the production rate of 47S rRNA precursor transcripts was dramatically reduced as an early event after drug treatment. Transcriptional inhibition of rRNA was followed by a robust G1 arrest, and activation of apoptotic proteins caspase-8, -9, and -3 and PARP-1 in a p53-independent manner. Using cell synchronization and flow cytometry, it was determined that cells treated while in G1 arrest immediately, but cells treated in S or G2 successfully complete mitosis. Twenty-four hours after treatment, the majority of cells finally arrest in G1, but nearly one-third contained highly compacted DNA; a distinct biological feature that cannot be associated with mitosis, senescence, or apoptosis. This unique effect mirrored the efficient condensation of tRNA and DNA in cell-free systems. The combination of DNA compaction and apoptosis by TriplatinNC treatment conferred striking activity in platinum-resistant and/or p53 mutant or null cell lines. Taken together, our results support that the biological activity of TriplatinNC reflects reduced metabolic deactivation (substitution-inert compound not reactive to sulfur nucleophiles), high cellular accumulation, and novel consequences of high-affinity noncovalent DNA binding, producing a new profile and a further shift in the structure-activity paradigms for antitumor complexes.
authors.affiliation: Department of Chemistry and ‡Massey Cancer Center, Virginia Commonwealth University , Richmond, Virginia 23284, United States.
authors.forename: Erica J
authors.initials: EJ
authors.lastname: Peterson
authors.forename: Vijay R
authors.initials: VR
authors.lastname: Menon
authors.forename: Laura
authors.initials: L
authors.lastname: Gatti
authors.forename: Ralph
authors.initials: R
authors.lastname: Kipping
authors.forename: Dilhara
authors.initials: D
authors.lastname: Dewasinghe
authors.forename: Paola
authors.initials: P
authors.lastname: Perego
authors.forename: Lawrence F
authors.initials: LF
authors.lastname: Povirk
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Farrell
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry*; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000187:drug effects*; D002474:Cell-Free System; D004247:DNA / Q000737:chemistry*; D005434:Flow Cytometry; D045325:HCT116 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D018613:Microscopy, Confocal; D008938:Mitosis; D009154:Mutation; D009944:Organoplatinum Compounds / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D010984:Platinum / Q000627:therapeutic use*; D012335:RNA, Ribosomal / Q000737:chemistry*; D012343:RNA, Transfer / Q000737:chemistry; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D009944:Organoplatinum Compounds; D010455:Peptides; D010710:Phosphates; D012335:RNA, Ribosomal; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C000602580:triplatinNC; C015462:polyarginine; D010984:Platinum; D004247:DNA; D012343:RNA, Transfer; D001616:beta-Galactosidase; D020169:Caspases; C114062:BBR 3464
keywords: BBR3464; DNA compaction; TriplatinNC; nucleolus; platinum; rRNA