Article | Published:

NFκB regulates p21 expression and controls DNA damage-induced leukemic differentiation

Oncogenevolume 37pages36473656 (2018) | Download Citation


DNA damage exposure is a major modifier of cell fate in both normal and cancer tissues. In response to DNA damage, myeloid leukemia cells activate a poorly understood terminal differentiation process. Here, we show that the NFκB pathway directly activates expression of the proliferation inhibitor p21 in response to DNA damage in myeloid leukemia cells. In order to understand the role of this unexpected regulatory event, we ablated the NFκB binding site we identified in the p21 promoter, using CRISPR/Cas9-mediated genome editing. We found that NFκB-mediated p21 activation controls DNA damage-induced myeloid differentiation. Our results uncover a p53-independent pathway for p21 activation involved in controlling hematopoietic cell fate.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    Grove CS, Vassiliou GS. Acute myeloid leukaemia: a paradigm for the clonal evolution of cancer? Dis Model Mech. 2014;7:941–51.

  2. 2.

    Bruserud O, Gjertsen BT, Huang T. Induction of differentiation and apoptosis—a possible strategy in the treatment of adult acute myelogenous leukemia. Oncologist. 2000;5:454–62.

  3. 3.

    Watts JM, Tallman MS. Acute promyelocytic leukemia: what is the new standard of care? Blood Rev. 2014;28:205–12.

  4. 4.

    Sherman MH, Bassing CH, Teitell MA. Regulation of cell differentiation by the DNA damage response. Trends Cell Biol. 2011;21:312–9.

  5. 5.

    Santos MA, Faryabi RB, Ergen AV, Day AM, Malhowski A, Canela A, et al. DNA-damage-induced differentiation of leukaemic cells as an anti-cancer barrier. Nature. 2014;514:107–11.

  6. 6.

    Nijnik A, Woodbine L, Marchetti C, Dawson S, Lambe T, Liu C, et al. DNA repair is limiting for haematopoietic stem cells during ageing. Nature. 2007;447:686–90.

  7. 7.

    Rossi DJ, Bryder D, Seita J, Nussenzweig A, Hoeijmakers J, Weissman IL. Deficiencies in DNA damage repair limit the function of haematopoietic stem cells with age. Nature. 2007;447:725–9.

  8. 8.

    Asada M, Yamada T, Fukumuro K, Mizutani S. p21Cip1/WAF1 is important for differentiation and survival of U937 cells. Leukemia. 1998;12:1944–50.

  9. 9.

    el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, et al. WAF1, a potential mediator of p53 tumor suppression. Cell. 1993;75:817–25.

  10. 10.

    Georgakilas AG, Martin OA, Bonner WM. p21: a two-faced genome guardian. Trends Mol Med. 2017;23:310–9.

  11. 11.

    Cheung KJ, Horsman DE, Gascoyne RD. The significance of TP53 in lymphoid malignancies: mutation prevalence, regulation, prognostic impact and potential as a therapeutic target. Br J Haematol. 2009;146:257–69.

  12. 12.

    Hou HA, Chou WC, Kuo YY, Liu CY, Lin LI, Tseng MH, et al. TP53 mutations in de novo acute myeloid leukemia patients: longitudinal follow-ups show the mutation is stable during disease evolution. Blood Cancer J. 2015;5:e331.

  13. 13.

    Melo MB, Ahmad NN, Lima CS, Pagnano KB, Bordin S, Lorand-Metze I, et al. Mutations in the p53 gene in acute myeloid leukemia patients correlate with poor prognosis. Hematology. 2002;7:13–9.

  14. 14.

    Rizzo MG, Zepparoni A, Cristofanelli B, Scardigli R, Crescenzi M, Blandino G, et al. Wt-p53 action in human leukaemia cell lines corresponding to different stages of differentiation. Br J Cancer. 1998;77:1429–38.

  15. 15.

    Sugimoto K, Toyoshima H, Sakai R, Miyagawa K, Hagiwara K, Ishikawa F, et al. Frequent mutations in the p53 gene in human myeloid leukemia cell lines. Blood. 1992;79:2378–83.

  16. 16.

    Wolf D, Rotter V. Major deletions in the gene encoding the p53 tumor antigen cause lack of p53 expression in HL-60 cells. Proc Natl Acad Sci USA. 1985;82:790–4.

  17. 17.

    Figarola JL, Weng Y, Lincoln C, Horne D, Rahbar S. Novel dichlorophenyl urea compounds inhibit proliferation of human leukemia HL-60 cells by inducing cell cycle arrest, differentiation and apoptosis. Invest New Drugs. 2012;30:1413–25.

  18. 18.

    Lee S, Zhou G, Clark T, Chen J, Rowley JD, Wang SM. The pattern of gene expression in human CD15+myeloid progenitor cells. Proc Natl Acad Sci USA. 2001;98:3340–5.

  19. 19.

    Hayden MS, Ghosh S. NF-kappa B, the first quarter-century: remarkable progress and outstanding questions. Genes Dev. 2012;26:203–34.

  20. 20.

    Tilstra JS, Clauson CL, Niedernhofer LJ, Robbins PD. NF-kappaB in aging and disease. Aging Dis. 2011;2:449–65.

  21. 21.

    Huang TT, Wuerzberger-Davis SM, Wu ZH, Miyamoto S. Sequential modification of NEMO/IKKgamma by SUMO-1 and ubiquitin mediates NF-kappaB activation by genotoxic stress. Cell. 2003;115:565–76.

  22. 22.

    Huang da W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 2009;4:44–57.

  23. 23.

    Cancer Genome Atlas Research N, Ley TJ, Miller C, Ding L, Raphael BJ, Mungall AJ, et al. Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia. N Engl J Med. 2013;368:2059–74.

  24. 24.

    Gao J, Aksoy BA, Dogrusoz U, Dresdner G, Gross B, Sumer SO. et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal. 2013;6:pl1.

  25. 25.

    Liu M, Lee MH, Cohen M, Bommakanti M, Freedman LP. Transcriptional activation of the Cdk inhibitor p21 by vitamin D3 leads to the induced differentiation of the myelomonocytic cell line U937. Genes Dev. 1996;10:142–53.

  26. 26.

    Pennington KN, Taylor JA, Bren GD, Paya CV. IkappaB kinase-dependent chronic activation of NF-kappaB is necessary for p21(WAF1/Cip1) inhibition of differentiation-induced apoptosis of monocytes. Mol Cell Biol. 2001;21:1930–41.

  27. 27.

    Galanos P, Vougas K, Walter D, Polyzos A, Maya-Mendoza A, Haagensen EJ, et al. Chronic p53-independent p21 expression causes genomic instability by deregulating replication licensing. Nat Cell Biol. 2016;18:777–89.

  28. 28.

    Price JG, Idoyaga J, Salmon H, Hogstad B, Bigarella CL, Ghaffari S, et al. CDKN1A regulates Langerhans cell survival and promotes Treg cell generation upon exposure to ionizing irradiation. Nat Immunol. 2015;16:1060–8.

  29. 29.

    Wang J, Sun Q, Morita Y, Jiang H, Gross A, Lechel A, et al. A differentiation checkpoint limits hematopoietic stem cell self-renewal in response to DNA damage. Cell. 2012;148:1001–14.

  30. 30.

    Gupta D, Shah HP, Malu K, Berliner N, Gaines P. Differentiation and characterization of myeloid cells. Curr Protoc Immunol. 2014;104:Unit 22F 25.

Download references


We would like to thank Wafik el-Deiry, Sinisa Dovat, David Claxton, Gregory Yochum, Sergei Grigoryev, James Broach, and the Penn State Flow Cytometry Core for materials, advice, and support. This work was supported by the Department of Defense (award CA140303) and the St. Baldrick Foundation.

Author information


  1. Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, 500 University Drive, Hershey, PA, 17033, USA

    • Claudia M. Nicolae
    • , Michael J. O’Connor
    • , Daniel Constantin
    •  & George-Lucian Moldovan


  1. Search for Claudia M. Nicolae in:

  2. Search for Michael J. O’Connor in:

  3. Search for Daniel Constantin in:

  4. Search for George-Lucian Moldovan in:

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to George-Lucian Moldovan.

Electronic supplementary material

About this article

Publication history