Abstract
Cellular senescence is considered as a crucial mechanism of tumor suppression that helps to prevent the growth of cells at risk for neoplastic transformation. In normal cells, cellular senescence induces an irreversible cell cycle arrest in response to telomere dysfunction, oncogene activation, genotoxic stress and a persistent DNA damage response (DDR). This process is accompanied by dramatic changes in cell morphology as well as in the activity of several signaling pathways. The senescent phenotype is multifaceted. In addition to an obligatory proliferation arrest, senescent cells manifest various senescence markers: mTOR-mediated hypertrophic growth (cell size increase), cell flattening, senescence-associated β galactosidase (SA-β gal) staining, expression of negative cell cycle regulators p53, p21Waf1 and p16Ink4a, specific chromatin reorganization including DNA segments with chromatin alterations reinforcing senescence (DNA-SCARS), senescence-associated secretory phenotype (SASP) and other features. Here, we describe the protocols that are used to study histone deacetylase inhibitor (HDACI)-induced cellular senescence in transformed cells with a special emphasis on the morphological features of senescence.
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Acknowledgments
We thank members of our laboratory Drs. Svetlana Svetlikova, Maria Abramova, Alexander Kukushkin, Svetlana Zubova, Tatiana Bykova, Vasily Romanov, and Elena Bukreeva for their constant support and help during preparation of this chapter. We are grateful to Sergei Gordeev for help in figure preparation and Ksenia Strunnikova for critical reading of manuscript. The work was supported by Russian Foundation for Basic Research (RFBR): No. 12-04-01393-a (VAP) and No. 10-04-01152 (TVP), Russian Academy of Sciences Program “Molecular and Cell Biology” and grant from the St. Petersburg State University, Contract № 1.37.122.2011.
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Pospelova, T.V., Chitikova, Z.V., Pospelov, V.A. (2013). An Integrated Approach for Monitoring Cell Senescence. In: Galluzzi, L., Vitale, I., Kepp, O., Kroemer, G. (eds) Cell Senescence. Methods in Molecular Biology, vol 965. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-239-1_26
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DOI: https://doi.org/10.1007/978-1-62703-239-1_26
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