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.

  • Letter
  • Published:

Oncogene-induced Nrf2 transcription promotes ROS detoxification and tumorigenesis

Abstract

Reactive oxygen species (ROS) are mutagenic and may thereby promote cancer1. Normally, ROS levels are tightly controlled by an inducible antioxidant program that responds to cellular stressors and is predominantly regulated by the transcription factor Nrf2 (also known as Nfe2l2) and its repressor protein Keap1 (refs 2–5). In contrast to the acute physiological regulation of Nrf2, in neoplasia there is evidence for increased basal activation of Nrf2. Indeed, somatic mutations that disrupt the Nrf2–Keap1 interaction to stabilize Nrf2 and increase the constitutive transcription of Nrf2 target genes were recently identified, indicating that enhanced ROS detoxification and additional Nrf2 functions may in fact be pro-tumorigenic6. Here, we investigated ROS metabolism in primary murine cells following the expression of endogenous oncogenic alleles of Kras, Braf and Myc, and found that ROS are actively suppressed by these oncogenes. K-RasG12D, B-RafV619E and MycERT2 each increased the transcription of Nrf2 to stably elevate the basal Nrf2 antioxidant program and thereby lower intracellular ROS and confer a more reduced intracellular environment. Oncogene-directed increased expression of Nrf2 is a new mechanism for the activation of the Nrf2 antioxidant program, and is evident in primary cells and tissues of mice expressing K-RasG12D and B-RafV619E, and in human pancreatic cancer. Furthermore, genetic targeting of the Nrf2 pathway impairs K-RasG12D-induced proliferation and tumorigenesis in vivo. Thus, the Nrf2 antioxidant and cellular detoxification program represents a previously unappreciated mediator of oncogenesis.

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: Physiological expression of oncogenes lowers ROS.
Figure 2: Physiological expression of oncogenes activates the Nrf2 antioxidant program.
Figure 3: Activation of Nrf2 by K-Ras G12D occurs via the Raf-MEK-ERK-Jun pathway.
Figure 4: Evidence for Nrf2 antioxidant program in pancreatic cancer.

Similar content being viewed by others

References

  1. Shibutani, S., Takeshita, M. & Grollman, A. P. Insertion of specific bases during DNA synthesis past the oxidation-damaged base 8-oxodG. Nature 349, 431–434 (1991)

    Article  ADS  CAS  Google Scholar 

  2. Itoh, K. et al. Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev. 13, 76–86 (1999)

    Article  CAS  Google Scholar 

  3. Wakabayashi, N. et al. Keap1-null mutation leads to postnatal lethality due to constitutive Nrf2 activation. Nature Genet. 35, 238–245 (2003)

    Article  CAS  Google Scholar 

  4. Nguyen, T., Nioi, P. & Pickett, C. B. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J. Biol. Chem. 284, 13291–13295 (2009)

    Article  CAS  Google Scholar 

  5. Venugopal, R. & Jaiswal, A. K. Nrf2 and Nrf1 in association with Jun proteins regulate antioxidant response element-mediated expression and coordinated induction of genes encoding detoxifying enzymes. Oncogene 17, 3145–3156 (1998)

    Article  CAS  Google Scholar 

  6. Hayes, J. D. & McMahon, M. NRF2 and KEAP1 mutations: permanent activation of an adaptive response in cancer. Trends Biochem. Sci. 34, 176–188 (2009)

    Article  CAS  Google Scholar 

  7. Tuveson, D. A. et al. Endogenous oncogenic K-rasG12D stimulates proliferation and widespread neoplastic and developmental defects. Cancer Cell 5, 375–387 (2004)

    Article  CAS  Google Scholar 

  8. Lee, A. C. et al. Ras proteins induce senescence by altering the intracellular levels of reactive oxygen species. J. Biol. Chem. 274, 7936–7940 (1999)

    Article  CAS  Google Scholar 

  9. Mitsushita, J., Lambeth, J. D. & Kamata, T. The superoxide-generating oxidase Nox1 is functionally required for Ras oncogene transformation. Cancer Res. 64, 3580–3585 (2004)

    Article  CAS  Google Scholar 

  10. Cakir, Y. & Ballinger, S. W. Reactive species-mediated regulation of cell signaling and the cell cycle: the role of MAPK. Antioxid. Redox Signal. 7, 726–740 (2005)

    Article  CAS  Google Scholar 

  11. Recktenwald, C. V., Kellner, R., Lichtenfels, R. & Seliger, B. Altered detoxification status and increased resistance to oxidative stress by K-ras transformation. Cancer Res. 68, 10086–10093 (2008)

    Article  CAS  Google Scholar 

  12. Irani, K. et al. Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science 275, 1649–1652 (1997)

    Article  CAS  Google Scholar 

  13. Tanaka, H. et al. E2F1 and c-Myc potentiate apoptosis through inhibition of NF-κB activity that facilitates MnSOD-mediated ROS elimination. Mol. Cell 9, 1017–1029 (2002)

    Article  CAS  Google Scholar 

  14. Murphy, D. J. et al. Distinct thresholds govern Myc’s biological output in vivo . Cancer Cell 14, 447–457 (2008)

    Article  CAS  Google Scholar 

  15. Birney, E. et al. Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project. Nature 447, 799–816 (2007)

    Article  ADS  CAS  Google Scholar 

  16. Karreth, F. A., DeNicola, G. M., Winter, S. P. & Tuveson, D. A. C-Raf inhibits MAPK activation and transformation by B-RafV600E . Mol. Cell 36, 477–486 (2009)

    Article  CAS  Google Scholar 

  17. Keum, Y. S. et al. Mechanism of action of sulforaphane: inhibition of p38 mitogen-activated protein kinase isoforms contributing to the induction of antioxidant response element-mediated heme oxygenase-1 in human hepatoma HepG2 cells. Cancer Res. 66, 8804–8813 (2006)

    Article  CAS  Google Scholar 

  18. Kwak, M. K., Itoh, K., Yamamoto, M. & Kensler, T. W. Enhanced expression of the transcription factor Nrf2 by cancer chemopreventive agents: role of antioxidant response element-like sequences in the nrf2 promoter. Mol. Cell. Biol. 22, 2883–2892 (2002)

    Article  CAS  Google Scholar 

  19. Hingorani, S. R. et al. Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell 4, 437–450 (2003)

    Article  CAS  Google Scholar 

  20. Jackson, E. L. et al. Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras . Genes Dev. 15, 3243–3248 (2001)

    Article  CAS  Google Scholar 

  21. Chan, K., Lu, R., Chang, J. C. & Kan, Y. W. NRF2, a member of the NFE2 family of transcription factors, is not essential for murine erythropoiesis, growth, and development. Proc. Natl Acad. Sci. USA 93, 13943–13948 (1996)

    Article  ADS  CAS  Google Scholar 

  22. Kawaguchi, Y. et al. The role of the transcriptional regulator Ptf1a in converting intestinal to pancreatic progenitors. Nature Genet. 32, 128–134 (2002)

    Article  CAS  Google Scholar 

  23. Park, J. H. et al. Evidence for the aldo-keto reductase pathway of polycyclic aromatic trans-dihydrodiol activation in human lung A549 cells. Proc. Natl Acad. Sci. USA 105, 6846–6851 (2008)

    Article  ADS  CAS  Google Scholar 

  24. Zhu, P., Oe, T. & Blair, I. A. Determination of cellular redox status by stable isotope dilution liquid chromatography/mass spectrometry analysis of glutathione and glutathione disulfide. Rapid Commun. Mass Spectrom. 22, 432–440 (2008)

    Article  ADS  CAS  Google Scholar 

  25. Carroll, J. S. et al. Chromosome-wide mapping of estrogen receptor binding reveals long-range regulation requiring the forkhead protein FoxA1. Cell 122, 33–43 (2005)

    Article  CAS  Google Scholar 

  26. He, X., Chen, M. G., Lin, G. X. & Ma, Q. Arsenic induces NAD(P)H-quinone oxidoreductase I by disrupting the Nrf2·Keap1·Cul3 complex and recruiting Nrf2·Maf to the antioxidant response element enhancer. J. Biol. Chem. 281, 23620–23631 (2006)

    Article  CAS  Google Scholar 

  27. Sun, J. et al. Hemoprotein Bach1 regulates enhancer availability of heme oxygenase-1 gene. EMBO J. 21, 5216–5224 (2002)

    Article  CAS  Google Scholar 

  28. Suvorova, E. S. et al. Cytoprotective Nrf2 pathway is induced in chronically txnrd 1-deficient hepatocytes. PLoS ONE 4, e6158 (2009)

    Article  ADS  Google Scholar 

  29. Rasband, W. S. ImageJ. (U. S. National Institutes of Health, 1997–2011) 〈http://imagej.nih.gov/ij/〉.

  30. Hahn, S. A. et al. Allelotype of pancreatic adenocarcinoma using xenograft enrichment. Cancer Res. 55, 4670–4675 (1995)

    CAS  PubMed  Google Scholar 

  31. Sun, C. et al. Characterization of the mutations of the K-ras, p53, p16, and SMAD4 genes in 15 human pancreatic cancer cell lines. Oncol. Rep. 8, 89–92 (2001)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank J. Johnson for providing the Nrf2−/− mice; G. Evan, O. Sansom, C. Murtaugh, J.-J. Ventura and E. Wagner for MEFs; E. Schmidt for Nrf2 antiserum; E. Jaffee, A. Maitra and A. Horii for human PDA cell lines; B. Haynes, S. Davies and N. Cook for human PDA tissue samples; C. Ross-Innes, K. Holmes and J. Carroll for advice with the ChIP assay; and the ENCODE Consortium for ChIP-seq studies. We thank F. Connor, C. Martins and other members of the Tuveson lab for assistance and advice, and the animal care staff and histology core at CRI. This research was supported by the University of Cambridge and Cancer Research UK, The Li Ka Shing Foundation and Hutchison Whampoa Limited, the NIHR Cambridge Biomedical Research Centre, and the NIH (grants CA101973, CA111294, CA084291 and CA105490 to D.A.T.; CA62924 and CA128920 to S.E.K. and C.I.-D.; and CA106610 to C.I.-D.). Additional support was obtained from the Abramson Cancer Center of the University of Pennsylvania Pilot Grant IRG 78-002-26 (D.A.T.), Emerald Foundation (E.S.C.), the Marjorie Kovler Fund (S.E.K.) and the Ruth L. Kirschstein National Research Service Award F32CA123887-01 (K.F.). We regret that many primary references have been omitted due to space limitations.

Author information

Authors and Affiliations

Authors

Contributions

G.M.D., F.A.K., T.J.H., A.G. and K.F. performed cell culture and mouse experiments. C.W., D.M., K.H.Y. and I.A.B. performed 8-oxo-dGuo and glutathione assays. C.J.Y., E.S.C., F.S., J.M.W., R.H.H., C.I.-D. and S.E.K. performed Nrf2 and Keap1 sequencing. G.M.D. and D.A.T. designed the study and wrote the manuscript, and all authors commented on it.

Corresponding author

Correspondence to David A. Tuveson.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-19 with legends, a Supplementary Discussion and additional references. (PDF 1619 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

DeNicola, G., Karreth, F., Humpton, T. et al. Oncogene-induced Nrf2 transcription promotes ROS detoxification and tumorigenesis. Nature 475, 106–109 (2011). https://doi.org/10.1038/nature10189

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10189

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing: Cancer

Sign up for the Nature Briefing: Cancer newsletter — what matters in cancer research, free to your inbox weekly.

Get what matters in cancer research, free to your inbox weekly. Sign up for Nature Briefing: Cancer