Skip to main content
SpringerLink
Log in

Radiosensitization of wildtype p53 cancer cells by the MDM2-inhibitor PXN727 is associated with altered heat shock protein 70 (Hsp70) levels

  • Original Paper
  • Published:
Cell Stress and Chaperones Aims and scope

Abstract

The oncoprotein MDM2 (murine double minute 2) is often overexpressed in human tumors and thereby attenuates the function of the tumor suppressor p53. In this study, we investigated the effects of the novel MDM2-inhibitor PXN727 on p53 activation, cell proliferation, cell cycle distribution and radiosensitivity. Since the localization of heat shock protein 70 (Hsp70) exerts different effects on radioresistance of tumor cells, we investigated the impact of PXN727 on intracellular, membrane, and secreted Hsp70 levels. We could show that PXN727 exerts its effects on wildtype p53 (HCT116 p53+/+, A549) but not p53 depleted (HCT116 p53−/−) or mutated (FaDu) tumor cells. PXN727 activates p53, induces the expression of p21, reduces the proportion of cells in the radioresistant S-phase and induces senescence. Radiosensitivity was significantly increased by PXN727 in HCT116 p53+/+ tumor cells. Furthermore, PXN727 causes a downregulation of Hsp70 membrane expression and an upregulated secretion of Hsp70 in wildtype p53 tumor cells. Our data suggest that re-activation of p53 by MDM2-inhibition modulates Hsp70 membrane expression and secretion which might contribute to the radiosensitizing effect of the MDM2-inhibitor PXN727.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Arya AK, El-Fert A, Devling T, Eccles RM, Aslam MA, Rubbi CP, Vlatkovic N, Fenwick J, Lloyd BH, Sibson DR, Jones TM, Boyd MT (2010) Nutlin-3, the small-molecule inhibitor of MDM2, promotes senescence and radiosensitises laryngeal carcinoma cells harbouring wild-type p53. Br J Cancer 103:186–195

    Article  PubMed  CAS  Google Scholar 

  • Cao C, Shinohara ET, Subhawong TK, Geng L, Woon KK, Albert JM, Hallahan DE, Lu B (2006) Radiosensitization of lung cancer by nutlin, an inhibitor of murine double minute 2. Mol Cancer Ther 5:411–417

    Article  PubMed  CAS  Google Scholar 

  • Gastpar R, Gehrmann M, Bausero MA, Asea A, Gross C, Schroeder JA, Multhoff G (2005) Heat shock protein 70 surface-positive tumor exosomes stimulate migratory and cytolytic activity of natural killer cells. Cancer Res 65:5238–5247

    Article  PubMed  CAS  Google Scholar 

  • Gehrmann M, Pfister K, Hutzler P, Gastpar R, Margulis B, Multhoff G (2002) Effects of antineoplastic agents on cytoplasmic and membrane-bound heat shock protein 70 (Hsp70) levels. Biol Chem 383:1715–1725

    PubMed  CAS  Google Scholar 

  • Gehrmann M, Schmetzer H, Eissner G, Haferlach T, Hiddemann W, Multhoff G (2003) Membrane-bound heat shock protein 70 (Hsp70) in acute myeloid leukemia: a tumor specific recognition structure for the cytolytic activity of autologous NK cells. Haematologica 88:474–476

    PubMed  Google Scholar 

  • Gehrmann M, Marienhagen J, Eichholtz-Wirth H, Fritz E, Ellwart J, Jaattela M, Zilch T, Multhoff G (2005) Dual function of membrane-bound heat shock protein 70 (Hsp70), Bag-4, and Hsp40: protection against radiation-induced effects and target structure for natural killer cells. Cell Death Differ 12:38–51

    Article  PubMed  CAS  Google Scholar 

  • Gudkov AV, Komarova EA (2003) The role of p53 in determining sensitivity to radiotherapy. Nat Rev Cancer 3:117–129

    Article  PubMed  CAS  Google Scholar 

  • Impicciatore G, Sancilio S, Miscia S, Di PR (2010) Nutlins and ionizing radiation in cancer therapy. Curr Pharm Des 16:1427–1442

    Article  PubMed  CAS  Google Scholar 

  • Jackson JG, Pant V, Li Q, Chang LL, Quintas-Cardama A, Garza D, Tavana O, Yang P, Manshouri T, Li Y, El-Naggar AK, Lozano G (2012) p53-mediated senescence impairs the apoptotic response to chemotherapy and clinical outcome in breast cancer. Cancer Cell 21:793–806

    Article  PubMed  CAS  Google Scholar 

  • Lehmann BD, McCubrey JA, Jefferson HS, Paine MS, Chappell WH, Terrian DM (2007) A dominant role for p53-dependent cellular senescence in radiosensitization of human prostate cancer cells. Cell Cycle 6:595–605

    Article  PubMed  CAS  Google Scholar 

  • Lehmann BD, Paine MS, Brooks AM, McCubrey JA, Renegar RH, Wang R, Terrian DM (2008) Senescence-associated exosome release from human prostate cancer cells. Cancer Res 68:7864–7871

    Article  PubMed  CAS  Google Scholar 

  • Michaelis M, Rothweiler F, Barth S, Cinatl J, van Rikxoort M, Loschmann N, Voges Y, Breitling R, von Deimling A, Rodel F, Weber K, Fehse B, Mack E, Stiewe T, Doerr HW, Speidel D, Cinatl J Jr (2011) Adaptation of cancer cells from different entities to the MDM2 inhibitor nutlin-3 results in the emergence of p53-mutated multi-drug-resistant cancer cells. Cell Death Dis 2:e243

    Article  PubMed  CAS  Google Scholar 

  • Michaelis M, Rothweiler F, Agha B, Barth S, Voges Y, Loschmann N, von Deimling A, Breitling R, Wilhelm DH, Rodel F, Speidel D, Cinatl J Jr (2012) Human neuroblastoma cells with acquired resistance to the p53 activator RITA retain functional p53 and sensitivity to other p53 activating agents. Cell Death Dis 3:e294

    Article  PubMed  CAS  Google Scholar 

  • Miyachi M, Kakazu N, Yagyu S, Katsumi Y, Tsubai-Shimizu S, Kikuchi K, Tsuchiya K, Iehara T, Hosoi H (2009) Restoration of p53 pathway by nutlin-3 induces cell cycle arrest and apoptosis in human rhabdomyosarcoma cells. Clin Cancer Res 15:4077–4084

    Article  PubMed  CAS  Google Scholar 

  • Multhoff G, Botzler C, Wiesnet M, Muller E, Meier T, Wilmanns W, Issels RD (1995) A stress-inducible 72-kDa heat-shock protein (HSP72) is expressed on the surface of human tumor cells, but not on normal cells. Int J Cancer 61:272–279

    Article  PubMed  CAS  Google Scholar 

  • Rothweiler U, Czarna A, Krajewski M, Ciombor J, Kalinski C, Khazak V, Ross G, Skobeleva N, Weber L, Holak TA (2008) Isoquinolin-1-one inhibitors of the MDM2-p53 interaction. Chem Med Chem 3:1118–1128

    PubMed  CAS  Google Scholar 

  • Schilling D, Gehrmann M, Steinem C, De MA, Pockley AG, Abend M, Molls M, Multhoff G (2009) Binding of heat shock protein 70 to extracellular phosphatidylserine promotes killing of normoxic and hypoxic tumor cells. FASEB J 23:2467–2477

    Article  PubMed  CAS  Google Scholar 

  • Shangary S, Wang S (2008) Targeting the MDM2-p53 interaction for cancer therapy. Clin Cancer Res 14:5318–5324

    Article  PubMed  CAS  Google Scholar 

  • Sherman M, Multhoff G (2007) Heat shock proteins in cancer. Ann N Y Acad Sci 1113:192–201

    Article  PubMed  CAS  Google Scholar 

  • Supiot S, Hill RP, Bristow RG (2008) Nutlin-3 radiosensitizes hypoxic prostate cancer cells independent of p53. Mol Cancer Ther 7:993–999

    Article  PubMed  CAS  Google Scholar 

  • Yu X, Harris SL, Levine AJ (2006) The regulation of exosome secretion: a novel function of the p53 protein. Cancer Res 66:4795–4801

    Article  PubMed  CAS  Google Scholar 

  • Yu X, Riley T, Levine AJ (2009) The regulation of the endosomal compartment by p53 the tumor suppressor gene. FEBS J 276:2201–2212

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

The authors would like to thank Michaela Wagner, Caroline Hagedorn and Nathalie Lippmann for excellent technical assistance, Peter Vaupel and Christine Bayer for helpful discussion and Khashayar Fakhrian for clinical advice. This work was supported by the Helmholtz Zentrum München (Clinical Cooperation Group - “Innate Immunity in Tumor Biology”), Deutsche Forschungsgemeinschaft (SFB 824/1; Cluster of Excellence 158: Munich-Centre of Advanced Photonics; INST 95/980-1 FUGG, Gulmay irradiation device), BMBF (MOBITUM, 01EZ0826; Kompetenzverbund Strahlenforschung, 03NUK007E; Leading-Edge Cluster m4—Personalized Medicine and Targeted Therapies, 01EX1021C).

Conflict of interest

None

Author information

Authors and Affiliations

  1. Department of Radiotherapy and Radiooncology, Klinikum rechts der Isar, Technische Universität München, Ismaninger Str. 22, 81675, Munich, Germany

    Daniela Schilling, Michael Düwel, Michael Molls & Gabriele Multhoff

  2. CCG-Innate Immunity in Tumor Biology, Helmholtz Center Munich, German Research Center for Environmental Health, Munich, Germany

    Daniela Schilling & Gabriele Multhoff

Authors
  1. Daniela Schilling
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael Düwel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael Molls
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gabriele Multhoff
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gabriele Multhoff.

Additional information

Daniela Schilling and Michael Düwel contributed equally.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schilling, D., Düwel, M., Molls, M. et al. Radiosensitization of wildtype p53 cancer cells by the MDM2-inhibitor PXN727 is associated with altered heat shock protein 70 (Hsp70) levels. Cell Stress and Chaperones 18, 183–191 (2013). https://doi.org/10.1007/s12192-012-0369-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12192-012-0369-8

Keywords

Search

Navigation