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Poly(ADP-ribose) synthesis: a useful parameter for identifying apoptotic cells

M Donzelli1, C Negri, A Mandarino

  • 1Istituto di Genetica Biochimica Evoluzionistica CNR, Pavia, Italy.

The Histochemical Journal
|February 18, 1998
PubMed
Summary
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This study analyzes apoptosis in HeLa cells induced by bleomycin or growth factor depletion. Apoptotic cells exhibit DNA fragmentation and cell cycle arrest, visualized through a novel multi-parameter assay.

Area of Science:

  • Cell Biology
  • Molecular Biology

Background:

  • Apoptosis is a crucial programmed cell death pathway.
  • Understanding apoptosis in cancer therapy and cellular stress is vital.

Purpose of the Study:

  • To analyze apoptosis in HeLa cells under different stress conditions.
  • To investigate the interference of apoptosis with cell cycle progression.
  • To characterize apoptotic features using a novel multi-parameter assay.

Main Methods:

  • HeLa cells were treated with bleomycin or growth factor depletion.
  • Flow cytometry was used to analyze cell cycle progression (propidium iodide, PCNA antibody).
  • A novel assay simultaneously analyzed chromatin condensation, DNA degradation, and poly(ADP-ribose) synthesis.

Related Experiment Videos

Main Results:

  • Bleomycin treatment caused G2/M phase arrest and subdiploid DNA content.
  • Growth factor depletion also led to subdiploid DNA content from all cell cycle phases.
  • The novel assay successfully visualized apoptotic features like condensed chromatin and fragmented DNA.

Conclusions:

  • Apoptosis induction by bleomycin or growth factor depletion affects cell cycle progression.
  • Subdiploid DNA content is a marker of apoptosis in HeLa cells.
  • The developed assay is effective for identifying diverse apoptotic features in single cells.