Induction of apoptosis by penta-O-galloyl-beta-D-glucose through activation of caspase-3 in human leukemia HL-60 cells

Eur J Pharmacol. 1999 Sep 24;381(2-3):171-83. doi: 10.1016/s0014-2999(99)00549-x.

Abstract

Penta-O-galloyl-beta-D-glucose is structurally related to (-)-epigallocatechin gallate and is isolated from hydrolyzed tannin. Penta-O-galloyl-beta-D-glucose can inhibit tumor promotion by teleocidin. We investigated the effects of penta-O-galloyl-beta-D-glucose and various tea polyphenols on cell viability in human leukemia HL-60 cells. In this study, we demonstrated that penta-O-galloyl-beta-D-glucose was able to induce apoptosis in a concentration- and time-dependent manner; however, other polyphenols were less effective. We further investigated the molecular mechanisms of penta-O-galloyl-beta-D-glucose-induced apoptosis. Treatment with penta-O-galloyl-beta-D-glucose caused induction of caspase-3/CPP32 activity in dose- and time-dependent manner, but not caspase-1 activity, and induced the degradation of poly-(ADP-ribose) polymerase. Pretreatment with acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO) and Z-Val-Ala-Asp-fluoromethyl-ketone (Z-VAD-FMK) inhibited penta-O-galloyl-beta-D-glucose-induced DNA fragmentation. Furthermore, treatment with penta-O-galloyl-beta-D-glucose (50 microM) caused a rapid loss of mitochondrial transmembrane potential, release of mitochondrial cytochrome c into cytosol, and subsequent induction of procaspase-9 processing. Our results indicate that penta-O-galloyl-beta-D-glucose allows caspase-activated deoxyribonuclease to enter the nucleus and degrade chromosomal DNA, and induces DFF-45 (DNA fragmentation factor) degradation. These results lead to a working hypothesis that penta-O-galloyl-beta-D-glucose-induced apoptosis is triggered by the release of cytochrome c into the cytosol, procaspase-9 processing, activation of caspase-3, degradation of poly-(ADP-ribose) polymerase, and DNA fragmentation caused by the caspase-activated deoxyribonuclease through the digestion of DFF-45. The induction of apoptosis by penta-O-galloyl-beta-D-glucose may provide a pivotal mechanism for its cancer chemopreventive action.

Publication types

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

MeSH terms

  • Acridine Orange
  • Apoptosis / drug effects*
  • Blotting, Western
  • Caspase 3
  • Caspases / metabolism*
  • Cell Division / drug effects
  • DNA Fragmentation
  • DNA, Neoplasm / biosynthesis
  • DNA, Neoplasm / isolation & purification
  • Dose-Response Relationship, Drug
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology*
  • Flow Cytometry
  • Fluorescent Dyes
  • HL-60 Cells
  • Humans
  • Hydrolyzable Tannins*
  • Membrane Potentials / drug effects
  • Mitochondria / drug effects
  • Sodium-Potassium-Exchanging ATPase / antagonists & inhibitors*
  • Subcellular Fractions / drug effects
  • Tannins / pharmacology*

Substances

  • DNA, Neoplasm
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Hydrolyzable Tannins
  • Tannins
  • beta-penta-O-galloyl-glucose
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • Sodium-Potassium-Exchanging ATPase
  • Acridine Orange