Clonogenic Assays to Detect Cell Fate in Mitotic Catastrophe

José Manuel Bravo-San Pedro, Oliver Kepp, Allan Sauvat, Santiago Rello-Varona, Guido Kroemer, Laura Senovilla

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    3 Citations (Scopus)

    Résumé

    Mitotic catastrophe (MC) is a cell death modality induced by DNA damage that involves the activation of cell cycle checkpoints such as the “DNA structure checkpoint” and “spindle assembly checkpoint” (SAC) leading to aberrant mitosis. Depending on the signal, MC can drive the cell to death or to senescence. The suppression of MC favors aneuploidy. Several cancer therapies, included microtubular poisons and radiations, trigger MC. The clonogenic assay has been used to study the capacity of single cells to proliferate and to generate macroscopic colonies and to evaluate the efficacy of anticancer drugs. Nevertheless, this method cannot analyze MC events. Here, we report an improved technique based on the use of human colon cancer HCT116 stable expressing histone H2B-GFP and DsRed-centrin proteins, allowing to determine the capacity of cells to proliferate, and to determine changes in the nucleus and centrosomes.

    langue originaleAnglais
    titreMethods in Molecular Biology
    EditeurHumana Press Inc.
    Pages227-239
    Nombre de pages13
    Les DOIs
    étatPublié - 1 janv. 2021

    Série de publications

    NomMethods in Molecular Biology
    Volume2267
    ISSN (imprimé)1064-3745
    ISSN (Electronique)1940-6029

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