title: An integrated approach for monitoring cell senescence.
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.
authors.affiliation: Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia.
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Pospelova
authors.forename: Zhanna V
authors.initials: ZV
authors.lastname: Chitikova
authors.forename: Valery A
authors.initials: VA
authors.lastname: Pospelov
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects / Q000235:genetics; D003584:Cytological Techniques / Q000379:methods*; D004801:Eosine Yellowish-(YS) / Q000378:metabolism; D044127:Epigenesis, Genetic / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D011905:Genes, ras / Q000235:genetics; D006416:Hematoxylin / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006657:Histones / Q000235:genetics; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D009857:Oncogenes / Q000235:genetics; D013194:Staining and Labeling; D014945:Wound Healing; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C468783:H2AX protein, human; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D008565:Membrane Proteins; D001616:beta-Galactosidase; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin