Enhancement of cisplatin-induced cytotoxicity by 7-hydroxystaurosporine (UCN-01), a new G2-checkpoint inhibitor.
DNA-damaging agents arrest cell cycle progression at either G1 or G2. A variety of agents such as caffeine have been shown to abrogate the DNA damage-dependent G2 checkpoint and enhance cytotoxicity. Unfortunately, this strategy has not enhanced therapeutic activity because adequate concentrations of these modulators are not tolerated in vivo. Here, using Chinese hamster ovary cell lines, we show that the potent protein kinase inhibitor 7-hydroxy-staurosporine (UCN-01) abrogates the G2 arrest induced by the DNA-damaging agent cisplatin. UCN-01 not only was effective at inducing mitosis when added to G2-arrested cells but also prevented cells from arresting in G2 when added to S-phase cells. Furthermore, UCN-01 did not cause premature mitosis of S-phase cells; rather, the cells progressed to G2 before undergoing mitosis. These effects were observed at noncytotoxic concentrations of UCN-01 that alone had no effect on cell cycle passage. Furthermore, the same concentrations of UCN-01 that resulted in abrogation of the cisplatin-induced G2 arrest also enhanced cisplatin-induced cytotoxicity, as determined by a colony formation assay. UCN-01 enhanced cisplatin cytotoxicity up to 60-fold and reduced by 3-fold the concentration of cisplatin required to kill 90% of the cells. The concentrations of UCN-01 required for this enhancement have been shown to be well tolerated in animal models, suggesting that this combination may represent an effective strategy for enhancing cisplatin-based chemotherapeutic regimens.
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Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Eastman
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D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D006224:Cricetinae; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D016195:G2 Phase / Q000187:drug effects*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D019311:Staurosporine / Q000031:analogs & derivatives
D000470:Alkaloids; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; C054852:7-hydroxystaurosporine; D011493:Protein Kinase C; D019311:Staurosporine; D002945:Cisplatin