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[TNF-induced apoptosis and necrosis in myeloleukemia cells HL-60 is regulated by reactive oxygen metabolites

E A Martynova1, E V Poddubskaia, E P Polosukhina

  • 1State Institute of Nutrition, RAMS, Ustinsky proezd 2/14, Moscow, 109240 Russia. eamartinova@lycos.com

Biomeditsinskaia Khimiia
|October 23, 2003
PubMed
Abstract

Insights

Tumor necrosis factor (TNF) induces apoptosis and necrosis through reactive oxygen metabolites, ceramide, and caspases. Necrosis may represent a disrupted apoptotic program, termed aponecrosis.

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Tumor necrosis factor (TNF) is a key ligand for death receptors, initiating apoptosis and necrosis.
  • Reactive oxygen metabolites, ceramide, and caspases are implicated as mediators of TNF-induced cell death.
  • The precise mechanistic interplay between these mediators and their regulation by cell cycle requires further elucidation.

Purpose of the Study:

  • To investigate the mechanistic relationship between TNF-induced apoptosis and necrosis.
  • To explore the role of ceramide synthesis and sphingomyelin breakdown in TNF signaling pathways.
  • To understand the regulation of TNF-induced cell death by cell cycle and specific inhibitors.

Main Methods:

  • Utilized TNF-alpha to induce apoptosis and necrosis in cells.
  • Employed inhibitors of ceramide synthase (fumonisin B1) and mitochondrial respiration.
  • Investigated the effects of exogenous bacterial sphingomyelinase (SMase) and denitrophenol (DNP).
  • Analyzed the role of TNF-alpha receptors R1 and R2 using monoclonal antibodies.

Main Results:

  • Executor caspase-3 activation is regulated during early stages of both TNF-induced apoptosis and necrosis.
  • TNF-induced cell damage, encompassing both apoptosis and necrosis, is cell cycle-dependent.
  • Exogenous SMase potentiates TNF-alpha-induced apoptosis, highlighting ceramide's role in this pathway.
  • Denitrophenol (DNP) inhibits TNF-induced necrosis and can initiate necrosis after SMase and TNF co-treatment.
  • Monoclonal antibodies to TNF-R1 and TNF-R2 induced higher levels of necrosis than TNF alone.

Conclusions:

  • Ceramide synthesis, sphingomyelin breakdown, caspase activation, and reactive oxygen metabolites are essential for TNF-alpha-induced apoptosis and necrosis.
  • These cell death pathways are potentially regulated by the cell cycle.
  • TNF-initiated necrosis may be a form of disrupted apoptosis, termed aponecrosis.

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