Peroxynitrite induces apoptosis of HL-60 cells by activation of a caspase-3 family protease

Am J Physiol. 1998 Apr;274(4):C855-60. doi: 10.1152/ajpcell.1998.274.4.C855.

Abstract

Apoptosis is an active process critical for the homeostasis of organisms. Enzymes of the caspase family are responsible for executing this process. We have previously shown that peroxynitrite (ONOO-), a biological product generated from the interaction of nitric oxide and superoxide, induces apoptosis of HL-60 cells. The aim of this study was to elucidate the mechanisms involved in the execution process of peroxynitrite-induced apoptosis. Proteolytic cleavage of poly(ADP-ribose) polymerase, an indication of caspase-3 family protease activation and an early biochemical event accompanying apoptosis, was observed in a time-dependent manner during peroxynitrite-induced apoptosis of HL-60 cells. Activation of caspase-3 during peroxynitrite-induced apoptosis was substantiated by monitoring proteolysis of the caspase-3 proenzyme and by measuring caspase-3 activity with a fluorogenic substrate. Furthermore, pretreatment of HL-60 cells with N-acetyl-Asp-Glu-Val-Asp-aldehyde, a specific inhibitor of caspase-3, but not N-acetyl-Tyr-Val-Ala-Asp-aldehyde, a specific inhibitor of caspase-1, decreased peroxynitrite-induced apoptosis. These results suggest that the activation of a caspase-3 family protease is essential for initiating the execution process of peroxynitrite-induced apoptosis of HL-60 cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Apoptosis* / drug effects
  • Apoptosis* / physiology
  • Caspase 3
  • Caspases*
  • Cysteine Endopeptidases / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology
  • Enzyme Activation / physiology
  • HL-60 Cells / drug effects*
  • HL-60 Cells / physiology
  • Humans
  • Nitrates / pharmacology*
  • Oxidants / pharmacology*
  • Peptide Hydrolases / metabolism
  • Poly(ADP-ribose) Polymerases / metabolism

Substances

  • Cysteine Proteinase Inhibitors
  • Nitrates
  • Oxidants
  • peroxynitric acid
  • Poly(ADP-ribose) Polymerases
  • Peptide Hydrolases
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • Cysteine Endopeptidases