Cellular senescence is the process of the permanent proliferative arrest of cells in response to various inducers. It is accompanied by typical morphological changes, in addition to the secretion of bioactive molecules, including proinflammatory cytokines and chemokines [known as the senescence-associated secretory phenotype (SASP)]. Thus, senescent cells may affect their local environment and induce a so-called 'bystander' senescence through the state of SASP. The phenotypes of senescent cells are determined by the type of agent inducing cellular stress and the cell lineages. To characterise the phenotypes of senescent cancer cells, two murine cell lines were employed in the present study: TC-1 and B16F10 (B16) cells. Two distinct senescence inductors were used: Chemotherapeutic agent docetaxel (DTX) and a combination of immunomodulatory cytokines, including interferon γ (IFNγ) and tumour necrosis factor α (TNFα). It was demonstrated that DTX induced senescence in TC-1 and B16 tumour cell lines, which was demonstrated by growth arrest, positive β-galactosidase staining, increased p21Waf1 (p21) expression and the typical SASP capable of inducing a 'bystander' senescence. By contrast, treatment with a combination of T helper cell 1 cytokines, IFNγ and TNFα, induced proliferation arrest only in B16 cells. Despite the presence of certain characteristic features resembling senescent cells (proliferation arrest, morphological changes and increased p21 expression), these cells were able to form tumours in vivo and started to proliferate upon cytokine withdrawal. In addition, B16 cells were not able to induce a 'bystander' senescence. In summary, the present study described cell line- and treatment-associated differences in the phenotypes of senescent cells that may be relevant in optimization of cancer chemo- and immunotherapy.

Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences v v i Prague 4 142 20 Czech Republic

Laboratory of Genome Integrity Institute of Molecular Genetics of the Czech Academy of Sciences v v i Prague 4 142 20 Czech Republic

Laboratory of Immunological and Tumour Models Institute of Molecular Genetics of the Czech Academy of Sciences v v i Prague 4 142 20 Czech Republic

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Coppé JP, Desprez PY, Krtolica A, Campisi J. The senescence-associated secretory phenotype: The dark side of tumor suppression. Annu Rev Pathol. 2010;5:99–118. doi: 10.1146/annurev-pathol-121808-102144. PubMed DOI PMC

Coppé JP, Patil CK, Rodier F, Sun Y, Muñoz DP, Goldstein J, Nelson PS, Desprez PY, Campisi J. Senescence-asso ciated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol. 2008;6:2853–2868. doi: 10.1371/journal.pbio.0060301. PubMed DOI PMC

Benvenuti S, Cramer R, Bruce J, Waterfield MD, Jat PS. Identification of novel candidates for replicative senescence by functional proteomics. Oncogene. 2002;21:4403–4413. doi: 10.1038/sj.onc.1205525. PubMed DOI

Bringold F, Serrano M. Tumor suppressors and oncogenes in cellular senescence. Exp Gerontol. 2000;35:317–329. doi: 10.1016/S0531-5565(00)00083-8. PubMed DOI

Cristofalo VJ, Lorenzini A, Allen RG, Torres C, Tresini M. Replicative senescence: A critical review. Mech Ageing Dev. 2004;125:827–848. doi: 10.1016/j.mad.2004.07.010. PubMed DOI

Salama R, Sadaie M, Hoare M, Narita M. Cellular senescence and its effector programs. Genes Dev. 2014;28:99–114. doi: 10.1101/gad.235184.113. PubMed DOI PMC

Pluquet O, Pourtier A, Abbadie C. The unfolded protein response and cellular senescence. A review in the theme: Cellular mechanisms of endoplasmic reticulum stress signaling in health and disease. Am J Physiol Cell Physiol. 2015;308:C415–C425. doi: 10.1152/ajpcell.00334.2014. PubMed DOI

Kojima H, Inoue T, Kunimoto H, Nakajima K. IL-6STAT3signaling and premature senescence. JAK-STAT. 2013;2:e25763. doi: 10.4161/jkst.25763. PubMed DOI PMC

Hayflick L, Moorhead PS. The serial cultivation of human diploid cell strains. Exp Cell Res. 1961;25:585–621. doi: 10.1016/0014-4827(61)90192-6. PubMed DOI

d’Adda di Fagagna F, Reaper PM, Clay-Farrace L, Fiegler H, Carr P, Von Zglinicki T, Saretzki G, Carter NP, Jackson SP. A DNA damage checkpoint response in telomere-initiated senescence. Nature. 2003;426:194–198. doi: 10.1038/nature02118. PubMed DOI

Harley CB, Futcher AB, Greider CW. Telomeres shorten during ageing of human fibroblasts. Nature. 1990;345:458–460. doi: 10.1038/345458a0. PubMed DOI

Blazkova H, Krejcikova K, Moudry P, Frisan T, Hodny Z, Bartek J. Bacterial intoxication evokes cellular senescence with persistent DNA damage and cytokine signalling. J Cell Mol Med. 2010;14:357–367. doi: 10.1111/j.1582-4934.2009.00862.x. PubMed DOI PMC

Cairney CJ, Bilsland AE, Evans TR, Roffey J, Bennett DC, Narita M, Torrance CJ, Keith WN. Cancer cell senescence: A new frontier in drug development. Drug Discov Today. 2012;17:269–276. doi: 10.1016/j.drudis.2012.01.019. PubMed DOI

Kuilman T, Michaloglou C, Mooi WJ, Peeper DS. The essence of senescence. Genes Dev. 2010;24:2463–2479. doi: 10.1101/gad.1971610. PubMed DOI PMC

Pascal T, Debacq-Chainiaux F, Chrétien A, Bastin C, Dabée AF, Bertholet V, Remacle J, Toussaint O. Comparison of replicative senescence and stress-induced premature senescence combining differential display and low-density DNA arrays. FEBS Lett. 2005;579:3651–3659. doi: 10.1016/j.febslet.2005.05.056. PubMed DOI

Davalos AR, Coppe JP, Campisi J, Desprez PY. Senescent cells as a source of inflammatory factors for tumor progression. Cancer Metastasis Rev. 2010;29:273–283. doi: 10.1007/s10555-010-9220-9. PubMed DOI PMC

Hubackova S, Krejcikova K, Bartek J, Hodny Z. IL1- and TGFβ-Nox4 signaling, oxidative stress and DNA damage response are shared features of replicative, oncogene-induced, and drug-induced paracrine ‘bystander senescence’. Aging (Albany NY) 2012;4:932–951. doi: 10.18632/aging.100520. PubMed DOI PMC

Calcinotto A, Alimonti A. Aging tumour cells to cure cancer: ‘pro-senescence’ therapy for cancer. Swiss Med Wkly. 2017;147:w14367. PubMed

Ortiz-Montero P, Londoño-Vallejo A, Vernot JP. Senescence- associated IL-6 and IL-8 cytokines induce a self- and cross-reinforced senescence/inflammatory milieu strengthening tumorigenic capabilities in the MCF-7 breast cancer cell line. Cell Commun Signal. 2017;15:17. doi: 10.1186/s12964-017-0172-3. PubMed DOI PMC

Rodier F, Campisi J. Four faces of cellular senescence. J Cell Biol. 2011;192:547–556. doi: 10.1083/jcb.201009094. PubMed DOI PMC

Nelson G, Wordsworth J, Wang C, Jurk D, Lawless C, Martin- Ruiz C, von Zglinicki T. A senescent cell bystander effect: Senescence-induced senescence. Aging Cell. 2012;11:345–349. doi: 10.1111/j.1474-9726.2012.00795.x. PubMed DOI PMC

Hodny Z, Hubackova S, Bartek J. Cytokines shape chemotherapy-induced and ‘bystander’ senescence. Aging (Albany NY) 2010;2:375–376. doi: 10.18632/aging.100171. PubMed DOI PMC

Kuilman T, Michaloglou C, Vredeveld LC, Douma S, van Doorn R, Desmet CJ, Aarden LA, Mooi WJ, Peeper DS. Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell. 2008;133:1019–1031. doi: 10.1016/j.cell.2008.03.039. PubMed DOI

Ozturk M, Arslan-Ergul A, Bagislar S, Senturk S, Yuzugullu H. Senescence and immortality in hepatocellular carcinoma. Cancer Lett. 2009;286:103–113. doi: 10.1016/j.canlet.2008.10.048. PubMed DOI

Koturbash I, Rugo RE, Hendricks CA, Loree J, Thibault B, Kutanzi K, Pogribny I, Yanch JC, Engelward BP, Kovalchuk O. Irradiation induces DNA damage and modulates epigenetic effectors in distant bystander tissue in vivo. Oncogene. 2006;25:4267–4275. doi: 10.1038/sj.onc.1209467. PubMed DOI

Zhou H, Randers-Pehrson G, Waldren CA, Vannais D, Hall EJ, Hei TK. Induction of a bystander mutagenic effect of alpha particles in mammalian cells. Proc Natl Acad Sci USA. 2000;97:2099–2104. doi: 10.1073/pnas.030420797. PubMed DOI PMC

Braumüller H, Wieder T, Brenner E, Aßmann S, Hahn M, Alkhaled M, Schilbach K, Essmann F, Kneilling M, Griessinger C, et al. T-helper-1-cell cytokines drive cancer into senescence. Nature. 2013;494:361–365. doi: 10.1038/nature11824. PubMed DOI

Simova J, Sapega O, Imrichova T, Stepanek I, Kyjacova L, Mikyskova R, Indrova M, Bieblova J, Bubenik J, Bartek J, et al. Tumor growth accelerated by chemotherapy-induced senescent cells is suppressed by treatment with IL-12 producing cellular vaccines. Oncotarget. 2016;7:54952–54964. doi: 10.18632/oncotarget.10712. PubMed DOI PMC

Lin KY, Guarnieri FG, Staveley-O’Carroll KF, Levitsky HI, August JT, Pardoll DM, Wu TC. Treatment of established tumors with a novel vaccine that enhances major histocompatibility class II presentation of tumor antigen. Cancer Res. 1996;56:21–26. PubMed

Overwijk WW, Restifo NP. B16 as a mouse model for human melanoma. Curr Protoc Immunol Chapter: 20 Unit. 2001;20:1. PubMed PMC

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25:402–408. doi: 10.1006/meth.2001.1262. PubMed DOI

Ramirez JM, Bai Q, Péquignot M, Becker F, Kassambara A, Bouin A, Kalatzis V, Dijon-Grinand M, De Vos J. Side scatter intensity is highly heterogeneous in undifferentiated pluripotent stem cells and predicts clonogenic self-renewal. Stem Cells Dev. 2013;22:1851–1860. doi: 10.1089/scd.2012.0658. PubMed DOI PMC

Zhang YH, Lin JX, Vilcek J. Interleukin-6 induction by tumor necrosis factor and interleukin-1 in human fibroblasts involves activation of a nuclear factor binding to a kappa B-like sequence. Mol Cell Biol. 1990;10:3818–3823. doi: 10.1128/MCB.10.7.3818. PubMed DOI PMC

Hubackova S, Kucerova A, Michlits G, Kyjacova L, Reinis M, Korolov O, Bartek J, Hodny Z. IFNγ induces oxidative stress, DNA damage and tumor cell senescence via TGFβ/SMAD signaling-dependent induction of Nox4 and suppression of ANT2. Oncogene. 2016;35:1236–1249. doi: 10.1038/onc.2015.162. PubMed DOI

Mukherjee S, Debata PR, Hussaini R, Chatterjee K, Baidoo JN, Sampat S, Szerszen A, Navarra JP, Fata J, Severinova E, et al. Unique synergistic formulation of curcumin, epicatechin gallate and resveratrol, tricurin, suppresses HPV E6, eliminates HPV+ cancer cells, and inhibits tumor progression. Oncotarget. 2017;8:60904–60916. PubMed PMC

Novakova Z, Hubackova S, Kosar M, Janderova-Rossmeislova L, Dobrovolna J, Vasicova P, Vancurova M, Horejsi Z, Hozak P, Bartek J, et al. Cytokine expression and signaling in drug- induced cellular senescence. Oncogene. 2010;29:273–284. doi: 10.1038/onc.2009.318. PubMed DOI

Ringel I, Horwitz SB. Studies with RP 56976 (taxotere): A semisynthetic analogue of taxol. J Natl Cancer Inst. 1991;83:288–291. doi: 10.1093/jnci/83.4.288. PubMed DOI

Schwarze SR, Fu VX, Desotelle JA, Kenowski ML, Jarrard DF. The identification of senescence-specific genes during the induction of senescence in prostate cancer cells. Neoplasia. 2005;7:816–823. doi: 10.1593/neo.05250. PubMed DOI PMC

Di Mitri D, Toso A, Chen JJ, Sarti M, Pinton S, Jost TR, D’Antuono R, Montani E, Garcia-Escudero R, Guccini I, et al. Tumour-infiltrating Gr-1+ myeloid cells antagonize senescence in cancer. Nature. 2014;515:134–137. doi: 10.1038/nature13638. PubMed DOI

Hernández-Vargas H, Palacios J, Moreno-Bueno G. Molecular profiling of docetaxel cytotoxicity in breast cancer cells: Uncoupling of aberrant mitosis and apoptosis. Oncogene. 2007;26:2902–2913. doi: 10.1038/sj.onc.1210102. PubMed DOI

Hernández-Vargas H, Palacios J, Moreno-Bueno G. Telling cells how to die: Docetaxel therapy in cancer cell lines. Cell Cycle. 2007;6:780–783. doi: 10.4161/cc.6.7.4050. PubMed DOI

Mikyšková R, Štěpánek I, Indrová M, Bieblová J, Šímová J, Truxová I, Moserová I, Fučíková J, Bartůňková J, Špíšek R, et al. Dendritic cells pulsed with tumor cells killed by high hydrostatic pressure induce strong immune responses and display therapeutic effects both in murine TC-1 and TRAMP-C2 tumors when combined with docetaxel chemotherapy. Int J Oncol. 2016;48:953–964. doi: 10.3892/ijo.2015.3314. PubMed DOI PMC

Kakimi K, Matsushita H, Hosoi A, Miyai M, Ohara O. CTLs regulate tumor growth via cytostatic effects rather than cytotoxicity: A few T cells can influence the growth of many times more tumor cells. OncoImmunology. 2014;4:e970464. doi: 10.4161/21624011.2014.970464. PubMed DOI PMC

Bartek J, Hodny Z, Lukas J. Cytokine loops driving senescence. Nat Cell Biol. 2008;10:887–889. doi: 10.1038/ncb0808-887. PubMed DOI

STAT3 inhibitor Stattic and its analogues inhibit STAT3 phosphorylation and modulate cytokine secretion in senescent tumour cells

Discovery of small molecule mechanistic target of rapamycin inhibitors as anti-aging and anti-cancer therapeutics

Synergism of BCL-2 family inhibitors facilitates selective elimination of senescent cells