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Flow Cytometric Analyses of p53-Mediated Cell Cycle Arrest and Apoptosis in Cancer Cells

Nour N Al Zouabi1, Cai M Roberts1, Z Ping Lin2

  • 1Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, USA.

Methods in Molecular Biology (Clifton, N.J.)
|May 25, 2021
PubMed

Insights

Studying gene effects like tumor suppressor p53 is challenging due to cell death. This method uses flow cytometry to quantify cell cycle and apoptosis, aiding long-term gene effect studies.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Stable gene expression can cause cell death, hindering phenotypic analysis.
  • Tumor suppressor p53 induces cell cycle arrest and apoptosis, but its long-term effects are hard to study.
  • Cellular context and damage level influence p53's pathway choice.

Purpose of the Study:

  • To describe a flow cytometric method for analyzing ectopic p53 expression.
  • To quantify cell cycle distribution and apoptosis in response to DNA damage.
  • To provide a method adaptable for studying other genes' effects on cell fate.

Main Methods:

  • Ectopic expression of p53 in cells.
  • Treatment of cells with DNA damaging agents.
  • Flow cytometric analysis of cell cycle distribution and apoptosis.

Main Results:

  • The method allows for quantification of cell cycle arrest and apoptosis.
  • It provides insights into the cellular response to p53 expression and DNA damage.
  • The technique is adaptable for various cell types and stress conditions.

Conclusions:

  • Flow cytometry offers a robust method for assessing gene expression impacts on cell cycle and apoptosis.
  • This approach facilitates the study of genes like p53 and their role in cellular stress responses.
  • The described method enhances the understanding of gene function and its contribution to cell fate determination.
Keywords:
ApoptosisCell cycle arrestDNA damageFlow cytometryGreen fluorescent proteinReporter plasmidp53

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