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Related Experiment Videos

Cell cycle-dependent tumor necrosis factor apoptosis

S C Shih1, O Stutman

  • 1The Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

Cancer Research
|April 1, 1996
PubMed
Summary

Tumor necrosis factor (TNF)-induced apoptosis in WEHI cells is linked to the cell cycle. These cells are most sensitive to TNF killing at the G1-S boundary, initiating apoptosis as they leave the S phase.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Immunology

Background:

  • Tumor necrosis factor (TNF) is a key mediator of apoptosis.
  • Understanding the cell cycle regulation of TNF-induced apoptosis is crucial for therapeutic development.

Purpose of the Study:

  • To investigate the relationship between TNF-mediated apoptosis and specific cell cycle stages in WEHI-164/2F cells.
  • To identify the precise cell cycle phase where WEHI cells are most susceptible to TNF-induced cell death.

Main Methods:

  • Cell synchronization at G0-G1 phase using low serum conditions.
  • Assessment of cell viability using MTT assays.
  • Quantification of apoptosis via propidium iodide staining and flow cytometry.
  • Analysis of DNA synthesis inhibition and cell cycle progression.

Main Results:

  • G0-G1 arrested WEHI cells exhibited increased sensitivity to TNF-induced apoptosis.
  • TNF inhibited DNA synthesis early in treatment and shifted cell cycle distribution towards S phase.
  • WEHI cells became progressively more sensitive to TNF as they approached the G1-S boundary.
  • TNF did not degrade DNA in cells that had passed the S phase before treatment.

Conclusions:

  • TNF-induced apoptosis in WEHI cells is cell cycle-dependent.
  • The G1-S boundary represents a critical window for WEHI cells to receive TNF cytotoxic signals.
  • Apoptosis is triggered as cells exit the S phase after TNF exposure.

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