Cytofluorometric assessment of acute cell death responses driven by radiation therapy

Beatriz Álvarez-Abril, Norma Bloy, Claudia Galassi, Ai Sato, Carlos Jiménez-Cortegana, Vanessa Klapp, Artur Aretz, Emma Guilbaud, Aitziber Buqué, Lorenzo Galluzzi*, Takahiro Yamazaki

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Radiation therapy (RT) is well known for its capacity to mediate cytostatic and cytotoxic effects on malignant cells, largely reflecting the ability of ionizing radiation to cause direct and indirect damage to macromolecules including DNA and lipids. While low-dose RT generally causes limited cytotoxicity in an acute manner (as it imposes insufficient cellular damage to compromise homeostasis, or instead induces the delayed demise of cells that fail to complete mitosis successfully), high RT doses can mediate an acute wave of cell death that begins to manifest shortly (24–72 h) after irradiation. Here, we provide two straightforward techniques to assess the acute cytotoxic effects of RT by the flow cytometry-assisted quantification of plasma membrane permeabilization (PMP, a late-stage manifestation of cell death) and either mitochondrial outer membrane permeabilization (MOMP) or phosphatidylserine (PS) externalization (two early-stage signs of cell death) in mouse mammary adenocarcinoma TS/A cells. With minor variations, the same protocols can be straightforwardly adapted to measure acute cell death responses as elicited by RT in a large panel of human and mouse cancer cells lines of different histological derivation.

Original languageEnglish
Title of host publicationRadiation Oncology and Radiotherapy Part A
EditorsAi Sato, Jeffrey Kraynak, Ariel E. Marciscano, Lorenzo Galluzzi
PublisherAcademic Press Inc.
Pages17-36
Number of pages20
ISBN (Print)9780323899499
DOIs
Publication statusPublished - Jan 2022
Externally publishedYes

Publication series

NameMethods in Cell Biology
Volume172
ISSN (Print)0091-679X

Keywords

  • Annexin V
  • Apoptosis
  • DiOC(3)
  • Mitochondrial outer membrane permeabilization
  • Plasma membrane permeabilization
  • Propidium iodide

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