Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Mouse double minute antagonist Nutlin-3a enhances chemotherapy-induced apoptosis in cancer cells with mutant p53 by activating E2F1

Abstract

MDM2 is a critical negative regulator of the p53 tumor suppressor protein. Recently, small-molecule antagonists of MDM2, the Nutlins, have been developed to inhibit the p53-MDM2 interaction and activate p53 signaling. However, half of human cancers have mutated p53 and they are resistant to Nutlin treatment. Here, we report that treatment of the p53-mutant malignant peripheral nerve sheath (MPNST) and p53-null HCT116 cells with cisplatin (Cis) and Nutlin-3a induced a degree of apoptosis that was significantly greater than either drug alone. Nutlin-3a also increased the cytotoxicity of both carboplatin and doxorubicin in a series of p53-mutant human tumor cell lines. In the human dedifferentiated liposarcoma cell line (LS141) and the p53 wild-type HCT116 cells, Nutlin-3a induced downregulation of E2F1 and this effect appeared to be proteasome dependent. In contrast, in MPNST and HCTp53−/− cells, Nutlin-3a inhibited the binding of E2F1 to MDM2 and induced transcriptional activation of free E2F1 in the presence of Cis-induced DNA damage. Downregulation of E2F1 by small interfering RNA significantly decreased the level of apoptosis induced by Cis and Nutlin-3a treatment. Moreover, expression of a dominant-negative form of E2F1 rescued cells from apoptosis, whereas cells overexpressing wild-type E2F1 showed an increase in cell death. This correlated with the induction of the proapoptotic proteins p73α and Noxa, which are both regulated by E2F1. These results indicate that antagonism of MDM2 by Nutlin-3a in cells with mutant p53 enhances chemosensitivity in an E2F1-dependent manner. Nutlin-3a therefore may provide a therapeutic benefit in tumors with mutant p53 provided it is combined with chemotherapy.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Banerjee D, Schnieders B, Fu JZ, Adhikari D, Zhao SC, Bertino JR . (1998). Role of E2F1 in chemosensitivity. Cancer Res 58: 4292–4296.

    CAS  PubMed  Google Scholar 

  • Bianco R, Ciardiello F, Tortora G . (2005). Chemosensitization by antisense oligonucleotides targeting MDM2. Curr Cancer Drug Targets 5: 51–56.

    Article  CAS  Google Scholar 

  • Bond GL, Hu W, Levine AJ . (2005). MDM2 is a central node in the p53 pathway: 12 years and counting. Curr Cancer Drug Targets 5: 3–8.

    Article  CAS  Google Scholar 

  • Carvajal D, Tovar C, Yang H, Vu BT, Heimbrook DC, Vassilev LT . (2005). Activation of p53 by MDM2 antagonists can protect proliferating cells from mitotic inhibitors. Cancer Res 65: 1918–1924.

    Article  CAS  Google Scholar 

  • Dai MS, Lu H . (2004). Inhibition of MDM2-mediated p53 ubiquitination and degradation by ribosomal protein L5. J Biol Chem 279: 44475–44482.

    Article  CAS  Google Scholar 

  • Furukawa Y, Nishimura N, Furukawa Y, Satoh M, Endo H, Iwase S et al. (2002). Apaf-1 is a mediator of E2F-1-induced apoptosis. J Biol Chem 277: 39760–39768.

    Article  CAS  Google Scholar 

  • Ganguli G, Wasylyk B . (2003). p53-independent functions of MDM2. Mol Cancer Res 1: 1027–1035.

    CAS  PubMed  Google Scholar 

  • Hershko T, Ginsberg D . (2004). Up-regulation of Bcl-2 homology 3 (BH3)-only proteins by E2F1 mediates apoptosis. J Biol Chem 279: 8627–8634.

    Article  CAS  Google Scholar 

  • Honda R, Yasuda H . (1999). Association of p19(ARF) with Mdm2 inhibits ubiquitin ligase activity of Mdm2 for tumor suppressor p53. EMBO J 18: 22–27.

    Article  CAS  Google Scholar 

  • Huang Y, Ishiko T, Nakada S, Utsugisawa T, Kato T, Yuan ZM . (1997). Role for E2F in DNA damage-induced entry of cells into S phase. Cancer Res 57: 3640–3643.

    CAS  PubMed  Google Scholar 

  • Irwin M, Marin MC, Phillips AC, Seelan RS, Smith DI, Liu W et al. (2000). Role for the p53 homologue p73 in E2F-1-induced apoptosis. Nature 407: 645–648.

    Article  CAS  Google Scholar 

  • Johnson DG, Schwarz JK, Cress WD, Nevins JR . (1993). Expression of transcription factor E2F1 induces quiescent cells to enter S phase. Nature 365: 349–352.

    Article  CAS  Google Scholar 

  • Kojima K, Konopleva M, Samudio IJ, Shikami M, Cabreira-Hansen M, McQueen T et al. (2005). MDM2 antagonists induce p53-dependent apoptosis in AML: implications for leukemia therapy. Blood 106: 3150–3159.

    Article  CAS  Google Scholar 

  • Kowalik TF, DeGregori J, Schwarz JK, Nevins JR . (1995). E2F1 overexpression in quiescent fibroblasts leads to induction of cellular DNA synthesis and apoptosis. J Virol 69: 2491–2500.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lin WC, Lin FT, Nevins JR . (2001). Selective induction of E2F1 in response to DNA damage, mediated by ATM-dependent phosphorylation. Genes Dev 15: 1833–1844.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Loughran O, La Thangue NB . (2000). Apoptotic and growth-promoting activity of E2F modulated by MDM2. Mol Cell Biol 20: 2186–2197.

    Article  CAS  Google Scholar 

  • Martin K, Trouche D, Hagemeier C, Sorensen TS, La Thangue NB, Kouzarides T . (1995). Stimulation of E2F1/DP1 transcriptional activity by MDM2 oncoprotein. Nature 375: 691–694.

    Article  CAS  Google Scholar 

  • Meng RD, Phillips P, El-Deiry WS . (1999). p53-independent increase in E2F-1 expression enhances the cytotoxic effects of etoposide and of adriamycin. Int J Oncol 14: 5–14.

    CAS  PubMed  Google Scholar 

  • Midgley CA, Lane DP . (1997). p53 protein stability in tumour cells is not determined by mutation but is dependent on Mdm2 binding. Oncogene 15: 1179–1189.

    Article  CAS  Google Scholar 

  • Momand J, Jung D, Wilczynski S, Niland J . (1998). The MDM2 gene amplification database. Nucleic Acids Res 26: 3453–3459.

    Article  CAS  Google Scholar 

  • Muller H, Bracken AP, Vernell R, Moroni MC, Christians F, Grassilli E et al. (2001). E2Fs regulate the expression of genes involved in differentiation, development, proliferation, and apoptosis. Genes Dev 15: 267–285.

    Article  CAS  Google Scholar 

  • Nip J, Strom DK, Fee BE, Zambetti G, Cleveland JL, Hiebert SW . (1997). E2F-1 cooperates with topoisomerase II inhibition and DNA damage to selectively augment p53-independent apoptosis. Mol Cell Biol 17: 1049–1056.

    Article  CAS  Google Scholar 

  • Pediconi N, Ianari A, Costanzo A, Belloni L, Gallo R, Cimino L et al. (2003). Differential regulation of E2F1 apoptotic target genes in response to DNA damage. Nat Cell Biol 5: 552–558.

    Article  CAS  Google Scholar 

  • Piette J, Neel H, Marechal V . (1997). Mdm2: keeping p53 under control. Oncogene 15: 1001–1010.

    Article  CAS  Google Scholar 

  • Qin XQ, Livingston DM, Kaelin Jr WG, Adams PD . (1994). Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis. Proc Natl Acad Sci USA 91: 10918–10922.

    Article  CAS  Google Scholar 

  • Stiewe T, Putzer BM . (2000). Role of the p53-homologue p73 in E2F1-induced apoptosis. Nat Genet 26: 464–469.

    Article  CAS  Google Scholar 

  • Stuhmer T, Chatterjee M, Hildebrandt M, Herrmann P, Gollasch H, Gerecke C et al. (2005). Nongenotoxic activation of the p53 pathway as a therapeutic strategy for multiple myeloma. Blood 106: 3609–3617.

    Article  Google Scholar 

  • Tovar C, Rosinski J, Filipovic Z, Higgins B, Kolinsky K, Hilton H et al. (2005). Small-molecule MDM2 antagonists reveal aberrant p53 signaling in cancer: implications for therapy. Proc Natl Acad Sci USA 103: 1888–1893.

    Article  Google Scholar 

  • Vassilev LT . (2005). p53 Activation by small molecules: application in oncology. J Med Chem 48: 4491–4499.

    Article  CAS  Google Scholar 

  • Vassilev LT, Vu BT, Graves B, Carvajal D, Podlaski F, Filipovic Z et al. (2004). In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science 303: 844–848.

    Article  CAS  Google Scholar 

  • Wasylyk C, Salvi R, Argentini M, Dureuil C, Delumeau I, Abecassis J et al. (1999). p53 mediated death of cells overexpressing MDM2 by an inhibitor of MDM2 interaction with p53. Oncogene 18: 1921–1934.

    Article  CAS  Google Scholar 

  • Wu X, Levine AJ . (1994). p53 and E2F-1 cooperate to mediate apoptosis. Proc Natl Acad Sci USA 91: 3602–3606.

    Article  CAS  Google Scholar 

  • Xiao ZX, Chen J, Levine AJ, Modjtahedi N, Xing J, Sellers WR et al. (1995). Interaction between the retinoblastoma protein and the oncoprotein MDM2. Nature 375: 694–698.

    Article  CAS  Google Scholar 

  • Yang Y, Ludwig RL, Jensen JP, Pierre SA, Medaglia MV, Davydov IV et al. (2005). Small molecule inhibitors of HDM2 ubiquitin ligase activity stabilize and activate p53 in cells. Cancer Cell 7: 547–559.

    Article  CAS  Google Scholar 

  • Zeng X, Chen L, Jost CA, Maya R, Keller D, Wang X et al. (1999). MDM2 suppresses p73 function without promoting p73 degradation. Mol Cell Biol 19: 3257–3266.

    Article  CAS  Google Scholar 

  • Zhang Z, Wang H, Li M, Rayburn ER, Agrawal S, Zhang R . (2005). Stabilization of E2F1 protein by MDM2 through the E2F1 ubiquitination pathway. Oncogene 24: 7238–7247.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to G K Schwartz.

Additional information

Supported in part by NEI Prostate Spore m. CA-92629.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ambrosini, G., Sambol, E., Carvajal, D. et al. Mouse double minute antagonist Nutlin-3a enhances chemotherapy-induced apoptosis in cancer cells with mutant p53 by activating E2F1. Oncogene 26, 3473–3481 (2007). https://doi.org/10.1038/sj.onc.1210136

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210136

Keywords

This article is cited by

Search

Quick links