Zinc Depletion by TPEN Induces Apoptosis in Human Acute Promyelocytic NB4 Cells

Cell Physiol Biochem. 2017;42(5):1822-1836. doi: 10.1159/000479539. Epub 2017 Jul 27.

Abstract

Background/aims: The effects of zinc signaling on proliferation or apoptosis of leukemia cells remain elusive. In the present study, we used N, N, N', N'-tetrakis-(2-pyridylmethyl)-ethylene-diamine (TPEN), a membrane-permeable zinc chelator, to evaluate the effect of zinc depletion on survival and apoptosis of NB4 acute promyelocytic leukemia (APL) cells.

Methods: The pro-apoptotic effects of TPEN on NB4 cells were examined by flow cytometry, and observed using an optical microscope. Intracellular labile zinc, nitric oxide (NO) or reactive oxygen species (ROS) changes caused by TPEN were measured by flow cytometry. We then explored possible roles of the crosstalk between intracellular labile zinc signaling and nitric oxide signaling in TPEN-triggered apoptosis.

Results: we found that TPEN induced apoptosis in NB4 APL cells in a dosage-dependent manner. We further demonstrated that TPEN triggered apoptosis by attenuating intracellular zinc and nitric oxide signaling in NB4 cells. Both exogenous zinc supplement and the nitric donor sodium nitroprusside (SNP) pre-incubation reversed TPEN-mediated inhibition of intracellular NO and Zn2+ signaling, and rescued NB4 cells from apoptosis.

Conclusion: These results suggest for the first time that crosstalk between zinc signaling and nitric oxide pathway is essential for the survival of NB4 cells. TPEN induces apoptosis in NB4 cells via negatively regulating intracellular NO and Zn2+ signaling. Our in vitro data suggest that zinc depletion by TPEN may be a potential therapeutic strategy for APL.

Keywords: Acute promyelocytic leukemia; Apoptosis; Nitric oxide; TPEN; Zinc.

MeSH terms

  • Apoptosis / drug effects*
  • Caspase Inhibitors / pharmacology
  • Caspases / chemistry
  • Caspases / metabolism
  • Cell Line, Tumor
  • Chelating Agents / chemistry
  • Chelating Agents / pharmacology*
  • Ethylenediamines / chemistry
  • Ethylenediamines / toxicity*
  • G1 Phase Cell Cycle Checkpoints / drug effects
  • Humans
  • Leukemia, Promyelocytic, Acute / metabolism
  • Leukemia, Promyelocytic, Acute / pathology
  • Nitric Oxide / metabolism
  • Nitroprusside / pharmacology
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / drug effects
  • Zinc / chemistry*
  • Zinc / metabolism

Substances

  • Caspase Inhibitors
  • Chelating Agents
  • Ethylenediamines
  • Reactive Oxygen Species
  • Nitroprusside
  • Nitric Oxide
  • Caspases
  • Zinc
  • N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine