title: M2, a novel anthracenedione, elicits a potent DNA damage response that can be subverted through checkpoint kinase inhibition to generate mitotic catastrophe.
abstract: Pixantrone is a promising anti-cancer aza-anthracenedione that has prompted the development of new anthracenediones incorporating symmetrical side-chains of increasing length varying from two to five methylene units in each pair of drug side-chains. A striking relationship has emerged in which anthracenedione-induced growth inhibition and apoptosis was inversely associated with side-chain length, a relationship that was attributable to a differential ability to stabilise the topoisomerase II (TOP2) cleavage complex. Processing of the complex to a DNA double strand break (DSB) flanked by γH2AX in nuclear foci is likely to occur, as the generation of the primary lesion was antecedent to γH2AX induction. M2, bearing the shortest pair of side-chains, induced TOP2-mediated DSBs efficiently and activated cell cycle checkpoints via Chk1 and Chk2 phosphorylation, implicating the involvement of ATM and ATR, and induced a protracted S phase and subsequent G2/M arrest. The inactive analogue M5, containing the longest pair of side-chains, only weakly stimulated any of these responses, suggesting that efficient stabilisation of the TOP2 cleavage complex was crucial for eliciting a strong DNA damage response (DDR). An M2 induced DDR in p53-defective MDA-MB-231 cells was abrogated by UCN-01, a ubiquitous inhibitor of kinases including Chk1, in a response associated with substantial mitotic catastrophe and strong synergy. The rational selection of checkpoint kinase inhibitors may significantly enhance the therapeutic benefit of anthracenediones that efficiently stabilise the TOP2 cleavage complex.
authors: Department of Biochemistry, La Trobe University, Victoria 3086, Australia.b.evison@latrobe.edu.au
authors: Benny J
authors: BJ
authors: Evison
authors: Mile
authors: M
authors: Pastuovic
authors: Rebecca A
authors: RA
authors: Bilardi
authors: Robert A
authors: RA
authors: Forrest
authors: Paul P
authors: PP
authors: Pumuye
authors: Brad E
authors: BE
authors: Sleebs
authors: Keith G
authors: KG
authors: Watson
authors: Don R
authors: DR
authors: Phillips
authors: Suzanne M
authors: SM
authors: Cutts
mesh_terms: D000818:Animals; D000880:Anthraquinones / Q000737:chemistry / Q000494:pharmacology*; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002417:Cattle; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D064447:Checkpoint Kinase 2; D004247:DNA / Q000737:chemistry; D004249:DNA Damage / Q000187:drug effects*; D004250:DNA Topoisomerases, Type II; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors; D004354:Drug Screening Assays, Antitumor; D004357:Drug Synergism; D006657:Histones / Q000096:biosynthesis; D006801:Humans; D008938:Mitosis / Q000187:drug effects*; D010539:Permeability; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D019311:Staurosporine / Q000031:analogs & derivatives / Q000494:pharmacology; D013329:Structure-Activity Relationship
chemical_list: D000880:Anthraquinones; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; C468783:H2AX protein, human; D006657:Histones; D000075223:Poly-ADP-Ribose Binding Proteins; D017382:Reactive Oxygen Species; C054852:7-hydroxystaurosporine; D004247:DNA; C427203:calf thymus DNA; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; C000605867:CHEK1 protein, human; C578634:CHEK2 protein, human; D000071877:Checkpoint Kinase 1; D017346:Protein Serine-Threonine Kinases; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human; D019311:Staurosporine