Assessment of cell cycle progression and mitotic slippage by videomicroscopy.

4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-08-21 DOI:10.1016/bs.mcb.2023.03.004
Luca Mattiello, Sara Soliman Abdel Rehim, Gwenola Manic, Ilio Vitale
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Abstract

Senescence is a state of irreversible cell cycle arrest accompanied by the acquisition of the senescence-associated secretory phenotype (SASP), which is activated in response to a variety of damaging stimuli, including genotoxic therapy. Accumulating evidence indicates that mitotic stress also promotes entry into senescence. This occurs via a mechanism involving defective mitoses and mitotic arrest, followed by abortion of cell division and slippage in the G1 phase. In this process, mitotic slippage leads to the generation of senescent cells characterized by a large cell body and a multinucleated and/or enlarged nuclear size. Here, we provide a detailed protocol for the assessment of cell proliferation and mitotic slippage in colorectal cancer cells upon pharmacological inhibition of the mitotic kinesin KIF11, best known as EG5. This approach can be used for preliminary characterization of senescence induction by therapeutics, but requires validation with standard senescence assays.

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通过视频显微镜评估细胞周期进展和有丝分裂滑动。
衰老是一种不可逆的细胞周期停滞状态,伴随着衰老相关分泌表型(SASP)的获得,SASP 会在包括基因毒性疗法在内的各种破坏性刺激下被激活。越来越多的证据表明,有丝分裂压力也会促进细胞进入衰老。其发生机制包括有丝分裂缺陷和有丝分裂停滞,随后是细胞分裂流产和 G1 期的滑动。在这一过程中,有丝分裂停止导致衰老细胞的产生,衰老细胞的特点是细胞体大、多核和/或核体积增大。在此,我们提供了一个详细的方案,用于评估药物抑制有丝分裂驱动蛋白 KIF11(又称 EG5)后结直肠癌细胞的增殖和有丝分裂滑动。这种方法可用于初步鉴定治疗药物的衰老诱导作用,但需要用标准衰老测定法进行验证。
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来源期刊
Methods in cell biology
Methods in cell biology 生物-细胞生物学
CiteScore
3.10
自引率
0.00%
发文量
125
审稿时长
3 months
期刊介绍: For over fifty years, Methods in Cell Biology has helped researchers answer the question "What method should I use to study this cell biology problem?" Edited by leaders in the field, each thematic volume provides proven, state-of-art techniques, along with relevant historical background and theory, to aid researchers in efficient design and effective implementation of experimental methodologies. Over its many years of publication, Methods in Cell Biology has built up a deep library of biological methods to study model developmental organisms, organelles and cell systems, as well as comprehensive coverage of microscopy and other analytical approaches.
期刊最新文献
Assessing chronological aging in Saccharomyces cerevisiae. Assessing microbiota composition in the context of aging. Assessing polyglutamine tract aggregation in the nematode Caenorhabditis elegans. Assessment of cell cycle progression and mitotic slippage by videomicroscopy. Cellular senescence and aging at the crossroad between immunity and cancer.
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