Modulation of G(2) arrest enhances cell death induced by the antitumor 1-nitroacridine derivative, Nitracrine.
Nitracrine (Ledakrin) is an antitumor drug which is activated by cellular enzymes and binds covalently to DNA. Previous studies have shown that covalent binding and crosslinking of DNA is associated with the cytotoxic and antitumor activities of this compound. In this study, cell cycle perturbations, effects on DNA synthesis and the cell death process initiated by Nitracrine were studied in murine leukemia L1210 cells. We show that exposure of L1210 cells to Nitracrine at the IC(99) concentration delayed progression through the S phase and transiently arrested cells in G(2)/M as found by flow cytometry. Higher drug concentration (2 x IC(99)) inhibited cell cycle progression in the S phase and induced rapid cell death. Both studied concentrations of the drug produced different effects on DNA synthesis as determined by bromodeoxyuridine incorporation, with a delay in the S phase progression at EC(99) concentration and irreversible arrest in early S phase at the higher dose (2 x IC(99)). At both concentrations of Nitracrine cell death occurred preferentially in the S phase as revealed by the TUNEL assay. When cells treated with the drug for 4 hours were post-incubated in the presence of 1 mM caffeine this led to rapid cell death and suppression of the G(2) arrest. This was associated with a about 10-fold increase in the cytotoxicity of Nitracrine. Similar effects were observed for another DNA crosslinking agent, cis-platinum, and to a lesser extent, for DNA topoisomerase I inhibitor, camptothecin. Together, our studies show that suppression of G(2) arrest induced by Nitracrine greatly enhances its cytotoxicity toward L1210 cells.
Skladanowski
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Department of Pharmaceutical Technology and Biochemistry, Technical University of Gdańsk, Narutowicza St. 11, 80-952 Gdańsk, Poland. as@altis.chem.pg.gda.pl
D017209:Apoptosis / Q000187:drug effects*; D002166:Camptothecin / Q000494:pharmacology; D002945:Cisplatin / Q000494:pharmacology; D053938:DNA Fragmentation; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D016195:G2 Phase / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D009564:Nitracrine / Q000494:pharmacology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D016196:S Phase / Q000187:drug effects
D019384:Nucleic Acid Synthesis Inhibitors; D009564:Nitracrine; D002945:Cisplatin; D002166:Camptothecin