CoChat
Sign Up
Cited by 181Telomeres, Telomerase, and SenescenceCancer-related Molecular PathwaysCell death mechanisms and regulation

Patterns of Early p21 Dynamics Determine Proliferation-Senescence Cell Fate after Chemotherapy

Open access
Hsu, Chien-Hsiang, Altschuler, Steven J., Wu, Lani F.·2019·Cell
DOI 10.1016/j.cell.2019.05.041PMC PMC6688629
Open this paper in CoChat — free accountOpen PDF

Abstract

Chemotherapy is designed to induce cell death. However, at non-lethal doses, cancer cells can choose to remain proliferative or become senescent. The slow development of senescence makes studying this decision challenging. Here, by analyzing single-cell p21 dynamics before, during, and days after drug treatment, we link three distinct patterns of early p21 dynamics to final cell fate. Surprisingly, while high p21 expression is classically associated with senescence, we find the opposite at early times during drug treatment: most senescence-fated cells express much lower p21 levels than proliferation-fated cells. We demonstrate that these dynamics lead to a p21 "Goldilocks zone" for proliferation, in which modest increases of p21 expression can lead to an undesirable increase of cancer cell proliferation. Our study identifies a counter-intuitive role for early p21 dynamics in the cell-fate decision and pinpoints a source of proliferative cancer cells that can emerge after exposure to non-lethal doses of chemotherapy.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.cell.2019.05.041

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Induction of p21 and a Notch-driven SASP are considered early events, whereas upregulation of p16INK4a and an NF-κB-driven SASP are evident at later stages of senescence.

    “Induction of p21 and a Notch- driven SASP are considered early events 70,75,76 , whereas upregulation of p16 INK4a and an NF- κB- driven SASP are evident at later stages of senescence 71,77,78 .”

    From Senescence and cancer — role and therapeutic opportunities (2022) · cites this paper as [76]

    The Results (page unavailable) report p21 induction at an early, 12-hour time point after chemotherapy, supporting the early-p21 part of the clause cited to this paper. They also find that most cells destined for senescence show delayed p21 induction, so the timing is not universal. Our copy does not discuss a Notch-driven SASP; the later-stage p16INK4a and NF-κB clause has a separate citation group. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The citation lists haven’t been fetched yet

References 84 · Cited by 181

Research works printed but not linked: 63

  1. Arora M, Moser J, Phadke H, Basha AA, and Spencer SL (2017). Endogenous Replication Stress in Mother Cells Leads to Quiescence of Daughter Cells. Cell Rep 19, 1351– 1364. 28514656 PMC5533606
  2. Banito A, and Lowe SW (2013). A new development in senescence. Cell 155, 977– 978. 24267881 PMC4702512
  3. Bashir T, Dorrello NV, Amador V, Guardavaccaro D, and Pagano M (2004). Control of the SCF(Skp2-Cks1) ubiquitin ligase by the APC/C(Cdh1) ubiquitin ligase. Nature 428, 190– 193. 15014502
  4. Beckerman R, Donner AJ, Mattia M, Peart MJ, Manley JL, Espinosa JM, and Prives C (2009). A role for Chk1 in blocking transcriptional elongation of p21 RNA during the S-phase checkpoint. Genes Dev 23, 1364– 1377. 19487575 PMC2701578
  5. Belloc F, Dumain P, Boisseau MR, Jalloustre C, Reiffers J, Bernard P, and Lacombe F (1994). A Flow Cytometric Method Using Hoechst-33342 and Propidium Iodide for Simultaneous Cell-Cycle Analysis and Apoptosis Determination in Unfixed Cells. Cytometry 17, 59– 65. 7528124
  6. Bieging KT, Mello SS, and Attardi LD (2014). Unravelling mechanisms of p53-mediated tumour suppression. Nat Rev Cancer 14, 359– 370. 24739573 PMC4049238
  7. Brown JP, Wei W, and Sedivy JM (1997). Bypass of senescence after disruption of p21CIP1/WAF1 gene in normal diploid human fibroblasts. Science 277, 831– 834. 9242615
  8. Burgess A, Chia KM, Haupt S, Thomas D, Haupt Y, and Lim E (2016). Clinical Overview of MDM2/X-Targeted Therapies. Front Oncol 6, 7. 26858935 PMC4728205
  9. Campisi J, and d’Adda di Fagagna F (2007). Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol 8, 729– 740. 17667954
  10. Cazzalini O, Scovassi AI, Savio M, Stivala LA, and Prosperi E (2010). Multiple roles of the cell cycle inhibitor p21(CDKN1A) in the DNA damage response. Mutat Res 704, 12– 20. 20096807
  11. Chang BD, Broude EV, Dokmanovic M, Zhu H, Ruth A, Xuan Y, Kandel ES, Lausch E, Christov K, and Roninson IB (1999). A senescence-like phenotype distinguishes tumor cells that undergo terminal proliferation arrest after exposure to anticancer agents. Cancer Res 59, 3761– 3767. 10446993
  12. Chang J, Wang Y, Shao L, Laberge RM, Demaria M, Campisi J, Janakiraman K, Sharpless NE, Ding S, Feng W, (2016). Clearance of senescent cells by ABT263 rejuvenates aged hematopoietic stem cells in mice. Nat Med 22, 78– 83. 26657143 PMC4762215
  13. Cooper S (2003). Rethinking synchronization of mammalian cells for cell cycle analysis. Cell Mol Life Sci 60, 1099– 1106. 12861378 PMC11138607
  14. Demaria M, O’Leary MN, Chang J, Shao L, Liu S, Alimirah F, Koenig K, Le C, Mitin N, Deal AM, (2017). Cellular Senescence Promotes Adverse Effects of Chemotherapy and Cancer Relapse. Cancer Discov 7, 165– 176. 27979832 PMC5296251
  15. Dimri GP, Lee XH, Basile G, Acosta M, Scott C, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereirasmith O, (1995). A Biomarker That Identifies Senescent Human-Cells in Culture and in Aging Skin in-Vivo. P Natl Acad Sci USA 92, 9363– 9367. PMC40985 7568133
  16. Eldeiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, and Vogelstein B (1993). Waf1, a Potential Mediator of P53 Tumor Suppression. Cell 75, 817– 825. 8242752
  17. Elmore LW, Di X, Dumur C, Holt SE, and Gewirtz DA (2005). Evasion of a single-step, chemotherapy-induced senescence in breast cancer cells: implications for treatment response. Clin Cancer Res 11, 2637– 2643. 15814644
  18. Ewald JA, Desotelle JA, Wilding G, and Jarrard DF (2010). Therapy-induced senescence in cancer. J Natl Cancer Inst 102, 1536– 1546. 20858887 PMC2957429
  19. Fang L, Igarashi M, Leung J, Sugrue MM, Lee SW, and Aaronson SA (1999). p21Waf1/Cip1/Sdi1 induces permanent growth arrest with markers of replicative senescence in human tumor cells lacking functional p53. Oncogene 18, 2789– 2797. 10362249
  20. Gewirtz DA (1999). A critical evaluation of the mechanisms of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunorubicin. Biochem Pharmacol 57, 727– 741. 10075079
  21. Greene RF, Collins JM, Jenkins JF, Speyer JL, and Myers CE (1983). Plasma pharmacokinetics of adriamycin and adriamycinol: implications for the design of in vitro experiments and treatment protocols. Cancer Res 43, 3417– 3421. 6850648
  22. He G, Siddik ZH, Huang Z, Wang R, Koomen J, Kobayashi R, Khokhar AR, and Kuang J (2005). Induction of p21 by p53 following DNA damage inhibits both Cdk4 and Cdk2 activities. Oncogene 24, 2929– 2943. 15735718
  23. Insinga A, Cicalese A, Faretta M, Gallo B, Albano L, Ronzoni S, Furia L, Viale A, and Pelicci PG (2013). DNA damage in stem cells activates p21, inhibits p53, and induces symmetric self-renewing divisions. Proc Natl Acad Sci U S A 110, 3931– 3936. 23417300 PMC3593901
  24. Janes KA, Albeck JG, Gaudet S, Sorger PK, Lauffenburger DA, and Yaffe MB (2005). A systems model of signaling identifies a molecular basis set for cytokine-induced apoptosis. Science 310, 1646– 1653. 16339439
  25. Jaqaman K, Loerke D, Mettlen M, Kuwata H, Grinstein S, Schmid SL, and Danuser G (2008). Robust single-particle tracking in live-cell time-lapse sequences. Nat Methods 5, 695– 702. 18641657 PMC2747604
  26. Jung YS, Qian Y, and Chen X (2010). Examination of the expanding pathways for the regulation of p21 expression and activity. Cell Signal 22, 1003– 1012. 20100570 PMC2860671
  27. Kang J, Hsu CH, Wu Q, Liu S, Coster AD, Posner BA, Altschuler SJ, and Wu LF (2016). Improving drug discovery with high-content phenotypic screens by systematic selection of reporter cell lines. Nat Biotechnol 34, 70– 77. 26655497 PMC4844861
  28. King C, Diaz H, Barnard D, Barda D, Clawson D, Blosser W, Cox K, Guo S, and Marshall M (2014). Characterization and preclinical development of LY2603618: a selective and potent Chk1 inhibitor. Invest New Drugs 32, 213– 226. 24114124
  29. Krenning L, Feringa FM, Shaltiel IA, van den Berg J, and Medema RH (2014). Transient activation of p53 in G2 phase is sufficient to induce senescence. Mol Cell 55, 59– 72. 24910099
  30. Kuilman T, Michaloglou C, Mooi WJ, and Peeper DS (2010). The essence of senescence. Genes Dev 24, 2463– 2479. 21078816 PMC2975923
  31. Kurz EU, Douglas P, and Lees-Miller SP (2004). Doxorubicin activates ATM-dependent phosphorylation of multiple downstream targets in part through the generation of reactive oxygen species. J Biol Chem 279, 53272– 53281. 15489221
  32. Kwon JS, Everetts NJ, Wang X, Wang WK, Della Croce K, Xing JH, and Yao G (2017). Controlling Depth of Cellular Quiescence by an Rb-E2F Network Switch. Cell Reports 20, 3223– 3235. 28954237 PMC6571029
  33. Lanni JS, and Jacks T (1998). Characterization of the p53-dependent postmitotic checkpoint following spindle disruption. Molecular and Cellular Biology 18, 1055– 1064. 9448003 PMC108818
  34. Ling YH, el-Naggar AK, Priebe W, and Perez-Soler R (1996). Cell cycle-dependent cytotoxicity, G2/M phase arrest, and disruption of p34cdc2/cyclin B1 activity induced by doxorubicin in synchronized P388 cells. Mol Pharmacol 49, 832– 841. 8622633
  35. Marechal A, and Zou L (2013). DNA damage sensing by the ATM and ATR kinases. Cold Spring Harb Perspect Biol 5. PMC3753707 24003211
  36. Nardella C, Clohessy JG, Alimonti A, and Pandolfi PP (2011). Pro-senescence therapy for cancer treatment. Nature Reviews Cancer 11, 503– 511. 21701512
  37. Nishitani H, Shiomi Y, Iida H, Michishita M, Takami T, and Tsurimoto T (2008). CDK inhibitor p21 is degraded by a proliferating cell nuclear antigen-coupled Cul4-DDB1Cdt2 pathway during S phase and after UV irradiation. J Biol Chem 283, 29045– 29052. 18703516 PMC2662008
  38. O’Connor MJ (2015). Targeting the DNA Damage Response in Cancer. Mol Cell 60, 547– 560. 26590714
  39. Overton KW, Spencer SL, Noderer WL, Meyer T, and Wang CL (2014). Basal p21 controls population heterogeneity in cycling and quiescent cell cycle states. Proc Natl Acad Sci U S A 111, E4386– 4393. 25267623 PMC4205626
  40. Paek AL, Liu JC, Loewer A, Forrester WC, and Lahav G (2016). Cell-to-Cell Variation in p53 Dynamics Leads to Fractional Killing. Cell 165, 631– 642. 27062928 PMC5217463
  41. Potter AJ, Gollahon KA, Palanca BJ, Harbert MJ, Choi YM, Moskovitz AH, Potter JD, and Rabinovitch PS (2002). Flow cytometric analysis of the cell cycle phase specificity of DNA damage induced by radiation, hydrogen peroxide and doxorubicin. Carcinogenesis 23, 389– 401. 11895853
  42. Rayess H, Wang MB, and Srivatsan ES (2012). Cellular senescence and tumor suppressor gene p16. Int J Cancer 130, 1715– 1725. 22025288 PMC3288293
  43. Reinhardt HC, Hasskamp P, Schmedding I, Morandell S, van Vugt MA, Wang X, Linding R, Ong SE, Weaver D, Carr SA, (2010). DNA damage activates a spatially distinct late cytoplasmic cell-cycle checkpoint network controlled by MK2-mediated RNA stabilization. Mol Cell 40, 34– 49. 20932473 PMC3030122
  44. Reyes J, Chen JY, Stewart-Ornstein J, Karhohs KW, Mock CS, and Lahav G (2018b). Fluctuations in p53 Signaling Allow Escape from Cell-Cycle Arrest. Molecular Cell 71, 581−+. PMC6282757 30057196
  45. Roninson IB (2003). Tumor cell senescence in cancer treatment. Cancer Res 63, 2705– 2715. 12782571
  46. Ruan S, Okcu MF, Ren JP, Chiao P, Andreeff M, Levin V, and Zhang W (1998). Overexpressed WAF1/Cip1 renders glioblastoma cells resistant to chemotherapy agents 1,3-bis(2-chloroethyl)-1-nitrosourea and cisplatin. Cancer Res 58, 1538– 1543. 9537261
  47. Ryl T, Kuchen EE, Bell E, Shao C, Florez AF, Monke G, Gogolin S, Friedrich M, Lamprecht F, Westermann F, (2017). Cell-Cycle Position of Single MYC-Driven Cancer Cells Dictates Their Susceptibility to a Chemotherapeutic Drug. Cell Syst 5, 237– 250 e238. 28843484
  48. Schmitt CA, Fridman JS, Yang M, Lee S, Baranov E, Hoffman RM, and Lowe SW (2002). A senescence program controlled by p53 and p16INK4a contributes to the outcome of cancer therapy. Cell 109, 335– 346. 12015983
  49. Seoane J, Le HV, and Massague J (2002). Myc suppression of the p21(Cip1) Cdk inhibitor influences the outcome of the p53 response to DNA damage. Nature 419, 729– 734. 12384701
  50. Shaltiel IA, Krenning L, Bruinsma W, and Medema RH (2015). The same, only different - DNA damage checkpoints and their reversal throughout the cell cycle. J Cell Sci 128, 607– 620. 25609713
  51. Shiotani B, and Zou L (2009). Single-stranded DNA orchestrates an ATM-to-ATR switch at DNA breaks. Mol Cell 33, 547– 558. 19285939 PMC2675165
  52. Smith J, Tho LM, Xu NH, and Gillespie DA (2010). The ATM-Chk2 and ATR-Chk1 Pathways in DNA Damage Signaling and Cancer. Advances in Cancer Research, Vol 108 108, 73– 112. 21034966
  53. Spencer SL, Cappell SD, Tsai FC, Overton KW, Wang CL, and Meyer T (2013). The Proliferation-Quiescence Decision Is Controlled by a Bifurcation in CDK2 Activity at Mitotic Exit. Cell 155, 369– 383. 24075009 PMC4001917
  54. Stewart-Ornstein J, and Lahav G (2016). Dynamics of CDKN1A in Single Cells Defined by an Endogenous Fluorescent Tagging Toolkit. Cell Rep 14, 1800– 1811. 26876176 PMC5154611
  55. Stoeger T, Battich N, Herrmann MD, Yakimovich Y, and Pelkmans L (2015). Computer vision for image-based transcriptomics. Methods 85, 44– 53. 26014038
  56. Suzuki K, Kostin S, Person V, Elsasser A, and Schaper J (2001). Time course of the apoptotic cascade and effects of caspase inhibitors in adult rat ventricular cardiomyocytes. J Mol Cell Cardiol 33, 983– 994. 11343420
  57. Taylor WR, and Stark GR (2001). Regulation of the G2/M transition by p53. Oncogene 20, 1803– 1815. 11313928
  58. te Poele RH, Okorokov AL, Jardine L, Cummings J, and Joel SP (2002). DNA damage is able to induce senescence in tumor cells in vitro and in vivo. Cancer Research 62, 1876– 1883. 11912168
  59. Uetake Y, and Sluder G (2010). Prolonged Prometaphase Blocks Daughter Cell Proliferation Despite Normal Completion of Mitosis. Curr Biol 20, 1666– 1671. 20832310 PMC2946429
  60. Wei W, Ayad NG, Wan Y, Zhang GJ, Kirschner MW, and Kaelin WG Jr. (2004). Degradation of the SCF component Skp2 in cell-cycle phase G1 by the anaphase-promoting complex. Nature 428, 194– 198. 15014503
  61. Xiao Z, Xue J, Sowin TJ, and Zhang H (2006). Differential roles of checkpoint kinase 1, checkpoint kinase 2, and mitogen-activated protein kinase-activated protein kinase 2 in mediating DNA damage-induced cell cycle arrest: implications for cancer therapy. Molecular cancer therapeutics 5, 1935– 1943. 16928813
  62. Yang F, Teves SS, Kemp CJ, and Henikoff S (2014). Doxorubicin, DNA torsion, and chromatin dynamics. Biochim Biophys Acta 1845, 84– 89. 24361676 PMC3927826
  63. Yang HW, Chung M, Kudo T, and Meyer T (2017). Competing memories of mitogen and p53 signalling control cell-cycle entry. Nature 549, 404– 408. 28869970 PMC6544019

Authors
Hsu, C.-H., Altschuler, S. J., & Wu, L. F.
Affiliations
University of California, San Francisco (US)
Corresponding author
  • Altschuler, Steven J.
  • Wu, Lani F.
ORCID
Venue
Cell
Publisher
Elsevier BV
Published
2019-07
Volume
178
Issue
2
Pages
361-373.e12
ISSN
0092-8674 (ISSN-L)
DOI
10.1016/j.cell.2019.05.041
PMC
PMC6688629
OpenAlex
W2950401027
Cited by
181
References
84
Citations by year
2026: 18 · 2025: 29 · 2024: 31 · 2023: 27 · 2022: 22 · 2021: 29 · 2020: 18 · 2019: 6
FWCI
12.86
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
National Institutes of Health
Topics
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Keywords
p21, chemotherapy, cellular senescence, tumor cell proliferation, cell fate determination, single-cell dynamics, habitable zone
MeSH terms
Checkpoint Kinase 1, DNA Damage, Doxorubicin, Humans, Models, Biological, Tumor Suppressor Protein p53, Cellular Senescence, RNA Interference, RNA, Small Interfering, Cell Line, Tumor, Cell Proliferation, Cyclin-Dependent Kinase Inhibitor p21, Cell Cycle Checkpoints

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review