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Using Sister Chromatid Exchange Assay to Detect Homologous Recombination Deficiency in Epigenetically Deregulated Urothelial Carcinoma Cells

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Urothelial Carcinoma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2684))

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Abstract

Sister chromatid exchange (SCE) is the process of exchanging regions between two sister chromatids during DNA replication. Exchanges between replicated chromatids and their sisters can be visualized in cells when DNA synthesis in one chromatid is labelled by 5-bromo-2’-deoxyuridine (BrdU). Homologous recombination (HR) is considered as the principal mechanism responsible for the sister chromatid exchange (SCE) upon replication fork collapse, and therefore SCE frequency upon genotoxic conditions reflects the capacity of HR repair to respond to replication stress. During tumorigenesis, inactivating mutations or altered transcriptome can affect a plethora of epigenetic factors that participate in DNA repair processes, and there are an increasing number of reports which demonstrate a link between epigenetic deregulation in cancer and homologous recombination deficiency (HRD). Therefore, the SCE assay can provide valuable information regarding the HR functionality in tumors with epigenetic deficiencies. In this chapter, we provide a method to visualize SCEs. The technique outlined below is characterized by high sensitivity and specificity and has been successfully applied to human bladder cancer cell lines. In this context, this technique could be used to characterize the dynamics of HR repair in tumors with deregulated epigenome.

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Acknowledgments

The authors apologize to all colleagues whose important contributions were not cited.

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Authors and Affiliations

  1. Biomedical Research Foundation of the Academy of Athens, Athens, Greece

    Theodoros Rampias & Apostolos Klinakis

Authors
  1. Theodoros Rampias
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  2. Apostolos Klinakis
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Corresponding authors

Correspondence to Theodoros Rampias or Apostolos Klinakis .

Editor information

Editors and Affiliations

  1. Department of Urology, Medical Faculty and University Hospital, Heinrich-Heine-University Duesseldorf, Düsseldorf, Germany

    Michèle J. Hoffmann

  2. Institute of Pathology, University Hospital, RWTH Aachen University, Aachen, Germany

    Nadine T. Gaisa

  3. Department of Urology, University Hospital Rechts der Isar, Technical University Munich, Munich, Germany

    Roman Nawroth

  4. Department of Urology, Helios Hospital, Bad Saarow, Germany

    Thorsten H. Ecke

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Rampias, T., Klinakis, A. (2023). Using Sister Chromatid Exchange Assay to Detect Homologous Recombination Deficiency in Epigenetically Deregulated Urothelial Carcinoma Cells. In: Hoffmann, M.J., Gaisa, N.T., Nawroth, R., Ecke, T.H. (eds) Urothelial Carcinoma. Methods in Molecular Biology, vol 2684. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3291-8_7

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  • DOI: https://doi.org/10.1007/978-1-0716-3291-8_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3290-1

  • Online ISBN: 978-1-0716-3291-8

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