Mechanism of dibucaine-induced apoptosis in promyelocytic leukemia cells (HL-60)

Kayo Arita, Toshihiko Utsumi, Akio Kato, Tomoko Kanno, Hirotsugu Kobuchi, Bunji Inoue, Jitsuo Akiyama, Kozo Utsumi

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58 Citations (Scopus)

Abstract

Dibucaine, a local anesthetic, inhibited the growth of promyelocytic leukemia cells (HL-60) without inducing arrest of the cell cycle and differentiation to granulocytes. Typical DNA fragmentation and DNA ladder formation were induced in a concentration- and time-dependent manner. The half-maximal concentration of dibucaine required to induce apoptosis was 100 μM. These effects were prevented completely by the pan-caspase inhibitor z- Val-Ala-Asp-(OMe)-fluoromethylketone (z-VAD-fmk), thereby implicating the cysteine aspartase (caspase) cascade in the process. Dibucaine activated various caspases, such as caspase-3, -6, -8, and -9 (-like) activities, but not caspase-1 (-like) activity, and induced mitochondrial membrane depolarization and the release of cytochrome c (Cyt.c) from mitochondria into the cytosol. Processing of pro-caspase-3, -8, and -9 by dibucaine was confirmed by western blot analysis. Bid, a death agonist member of the Bcl-2 family, was processed by caspases following exposure of cells to dibucaine. However, 100 μM dibucaine scarcely inhibited oxidative phosphorylation, but it induced membrane permeability transition in isolated rat liver mitochondria. Taken together, these data suggest that dibucaine induced apoptosis of HL-60 cells through activation of the caspase cascade in conjunction with Cyt.c release induced by a processed product of Bid and depolarization of the mitochondrial membrane potential. (C) 2000 Elsevier Science Inc.

Original languageEnglish
Pages (from-to)905-915
Number of pages11
JournalBiochemical Pharmacology
Volume60
Issue number7
DOIs
Publication statusPublished - Oct 1 2000
Externally publishedYes

Keywords

  • Apoptosis
  • Bid
  • Caspase activation
  • Cytochrome c
  • Dibucaine
  • Membrane permeability transition

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

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  • Cite this

    Arita, K., Utsumi, T., Kato, A., Kanno, T., Kobuchi, H., Inoue, B., Akiyama, J., & Utsumi, K. (2000). Mechanism of dibucaine-induced apoptosis in promyelocytic leukemia cells (HL-60). Biochemical Pharmacology, 60(7), 905-915. https://doi.org/10.1016/S0006-2952(00)00406-8