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JNK activation is required for TNFα-induced apoptosis in human hepatocarcinoma cells

Valerio Giacomo Minero1, Amina Khadjavi, Paola Costelli

  • 1Department of Experimental Medicine and Oncology, University of Turin, Italy.

International Immunopharmacology
|June 12, 2013
PubMed
Abstract

Insights

Tumor necrosis factor-alpha (TNF) combined with cycloheximide (CHX) induces apoptosis in hepatoma cells. This cell death involves both JNK and caspase pathways, offering targets to overcome cancer resistance.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Cancer Research

Background:

  • Hepatocellular carcinomas exhibit resistance to apoptosis induced by agents like tumor necrosis factor-alpha (TNF).
  • Hepatocytes are less susceptible to TNF-induced apoptosis due to NF-κB activation, but this can be overcome with protein synthesis inhibitors.

Purpose of the Study:

  • To investigate the molecular mechanisms of TNF and cycloheximide (CHX)-induced apoptosis in human hepatoma Huh7 cells.
  • To elucidate the specific role of JNK signaling in this apoptotic process.

Main Methods:

  • Huh7 cells were treated with TNF + CHX, with or without JNK inhibitor (SP600125) or pancaspase inhibitor (zVADfmk).
  • JNK expression was suppressed using RNA interference (RNAi).
  • Apoptosis, JNK activation, and caspase activation were assessed via microscopy, flow cytometry, western blotting, and enzymatic assays.

Main Results:

  • TNF + CHX-induced cell death was partially inhibited by blocking JNK activity or expression, indicating JNK's involvement.
  • The pancaspase inhibitor zVADfmk significantly reduced apoptosis.
  • Combined inhibition of JNK and caspases completely abrogated cell death.

Conclusions:

  • JNK and caspases play a causal role in TNF + CHX-induced apoptosis in Huh7 hepatoma cells.
  • Targeting both JNK and caspase pathways presents a promising strategy to overcome resistance to TNF-induced apoptosis in hepatocarcinoma.
Keywords:
ApoptosisCaspasesHepatocarcinomaJNKTNF

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