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Assessment of transcription inhibition as a characteristic of immunogenic cell death

Giulia Cerrato1, Juliette Humeau2, Allan Sauvat2

  • 1Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université, INSERM UMR1138, Centre de Recherche des Cordeliers, Paris, France; Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France; Université Paris Sud, Paris Saclay, Faculty of Medicine, Kremlin Bicêtre, Paris, France.

Methods in Cell Biology
|September 5, 2022
PubMed

Insights

Anticancer drugs inhibiting DNA-to-RNA transcription effectively trigger antitumor immune responses by stimulating immunogenic cell death. Precise measurement of transcription inhibition is crucial for this therapeutic approach.

Area of Science:

  • Molecular Biology
  • Immunology
  • Pharmacology

Background:

  • Anticancer drugs suppressing DNA-to-RNA transcription are effective in stimulating immunogenic cell death.
  • This mechanism eradicates malignant cells and ignites an antitumor immune response, leading to lasting therapeutic effects.
  • Precise measurement of transcription inhibition is essential for optimizing this treatment strategy.

Purpose of the Study:

  • To detail two complementary assays for accurately assessing transcription inhibition.
  • To provide reliable methods for quantifying the impact of anticancer drugs on RNA transcription.

Main Methods:

  • Two-color immunofluorescence assay to detect the physical separation of fibrillarin and nucleolin.
  • Click chemistry assay to measure the diminution of incorporated 5-ethynyl uridine (EU) into RNA, assessing per-cell fluorescence intensity.

Main Results:

  • The study presents two validated assays for measuring transcription inhibition.
  • These methods allow for precise quantification of drug-induced transcription suppression.

Conclusions:

  • Accurate assessment of transcription inhibition is vital for anticancer therapies that leverage immunogenic cell death.
  • The described assays provide valuable tools for evaluating drug efficacy and optimizing treatment strategies.
Keywords:
Cell stressCellular imagingClick chemistryDactinomycinRNA synthesis

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