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Updated: May 15, 2026
Simultaneous Imaging and Flow-Cytometry-based Detection of Multiple Fluorescent Senescence Markers in Therapy-Induced Senescent Cancer Cells
Published on: July 12, 2022
An integrated approach for monitoring cell senescence
Tatiana V Pospelova1, Zhanna V Chitikova, Valery A Pospelov
1Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia.
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, p21(Waf1) and p16(Ink4a), 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.
Insights
Histone deacetylase inhibitor-induced cellular senescence is a key tumor suppression mechanism. This study details protocols for observing the morphological hallmarks of senescence in transformed cells.
Area of Science:
- Oncology
- Cell Biology
- Molecular Biology
Background:
- Cellular senescence is a vital tumor suppression mechanism preventing neoplastic transformation.
- Senescence involves irreversible cell cycle arrest due to stressors like DNA damage.
- The senescent phenotype includes morphological changes and specific molecular markers.
Purpose of the Study:
- To describe protocols for studying histone deacetylase inhibitor (HDACI)-induced cellular senescence.
- To emphasize the morphological characteristics of HDACI-induced senescence in transformed cells.
Main Methods:
- Utilizing histone deacetylase inhibitors (HDACls) to induce cellular senescence.
- Focusing on morphological assessments of senescent cells.
- Observing key senescence markers such as cell size, shape, and SA-β gal staining.
Main Results:
- HDACI treatment reliably induces a senescent phenotype in transformed cells.
- Distinct morphological changes, including cell enlargement and flattening, are characteristic of HDACI-induced senescence.
- Specific senescence markers like SA-β gal are consistently observed.
Conclusions:
- HDACI-induced cellular senescence provides a valuable model for studying tumor suppression.
- Detailed morphological analysis is crucial for identifying and characterizing senescent cells.
- These protocols facilitate research into the role of senescence in cancer prevention.
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