Abstract
The Notch1 gene has an important role in mammalian cell-fate decision and tumorigenesis. Upstream control mechanisms for transcription of this gene are still poorly understood. In a chemical genetics screen for small molecule activators of Notch signalling, we identified epidermal growth factor receptor (EGFR) as a key negative regulator of Notch1 gene expression in primary human keratinocytes, intact epidermis and skin squamous cell carcinomas (SCCs). The underlying mechanism for negative control of the Notch1 gene in human cells, as well as in a mouse model of EGFR-dependent skin carcinogenesis, involves transcriptional suppression of p53 by the EGFR effector c-Jun. Suppression of Notch signalling in cancer cells counteracts the differentiation-inducing effects of EGFR inhibitors while, at the same time, synergizing with these compounds in induction of apoptosis. Thus, our data reveal a key role of EGFR signalling in the negative regulation of Notch1 gene transcription, of potential relevance for combinatory approaches for cancer therapy.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
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
Similar content being viewed by others
References
Lefort, K. & Dotto, G. P. Notch signaling in the integrated control of keratinocyte growth/differentiation and tumor suppression. Semin. Cancer Biol. 14, 374–386 (2004).
Lefort, K. et al. Notch1 is a p53 target gene involved in human keratinocyte tumor suppression through negative regulation of ROCK1/2 and MRCKα kinases. Genes Dev. 21, 562–577 (2007).
Artavanis-Tsakonas, S., Rand, M. D. & Lake, R. J. Notch signaling: cell fate control and signal integration in development. Science 284, 770–776 (1999).
Bray, S. J. Notch signalling: a simple pathway becomes complex. Nature Rev. Mol. Cell Biol. 7, 678–689 (2006).
Yugawa, T. et al. Regulation of Notch1 gene expression by p53 in epithelial cells. Mol. Cell Biol. 27, 3732–3742 (2007).
Scaltriti, M. & Baselga, J. The epidermal growth factor receptor pathway: a model for targeted therapy. Clin. Cancer Res. 12, 5268–5272 (2006).
Jost, M., Kari, C. & Rodeck, U. The EGF receptor — an essential regulator of multiple epidermal functions. Eur. J. Dermatol. 10, 505–510 (2000).
Zenz, R. & Wagner, E. F. Jun signalling in the epidermis: From developmental defects to psoriasis and skin tumors. Int. J. Biochem. Cell Biol. 38, 1043–1049 (2006).
Kalyankrishna, S. & Grandis, J. R. Epidermal growth factor receptor biology in head and neck cancer. J. Clin. Oncol. 24, 2666–2672 (2006).
Citri, A. & Yarden, Y. EGF–ERBB signalling: towards the systems level. Nature Rev. Mol. Cell Biol. 7, 505–516 (2006).
Lacouture, M. E. Mechanisms of cutaneous toxicities to EGFR inhibitors. Nature Rev. Cancer 6, 803–812 (2006).
Collins, I. & Workman, P. New approaches to molecular cancer therapeutics. Nature Chem. Biol. 2, 689–700 (2006).
Gong, Y. et al. Induction of BIM is essential for apoptosis triggered by EGFR kinase inhibitors in mutant EGFR-dependent lung adenocarcinomas. PLoS Med. 4, e294 (2007).
Scholl, F. A., Dumesic, P. A. & Khavari, P. A. Mek1 alters epidermal growth and differentiation. Cancer Res. 64, 6035–6040 (2004).
Friday, B. B. & Adjei, A. A. Advances in targeting the Ras/Raf/MEK/Erk mitogen-activated protein kinase cascade with MEK inhibitors for cancer therapy. Clin. Cancer Res. 14, 342–346 (2008).
Mandinova A. et al. The FoxO3A gene is a key negative target of canonical Notch signaling in the keratinocyte UVB response. EMBO J. 27, 1243–1254 (2008).
Laptenko, O. & Prives, C. Transcriptional regulation by p53: one protein, many possibilities. Cell Death Differ. 13, 951–961 (2006).
Vassilev, L. T. et al. In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science 303, 844–848 (2004).
Boggs, K. & Reisman, D. C/EBPβ participates in regulating transcription of the p53 gene in response to mitogen stimulation. J. Biol. Chem. 282, 7982–7990 (2007).
Bruno, T. et al. Che-1 phosphorylation by ATM/ATR and Chk2 kinases activates p53 transcription and the G2/M checkpoint. Cancer Cell 10, 473–486 (2006).
Phan, R. T. & Dalla-Favera, R. The BCL6 proto-oncogene suppresses p53 expression in germinal-centre B cells. Nature 432, 635–639 (2004).
Schreiber, M. et al. Control of cell cycle progression by c-Jun is p53 dependent. Genes Dev. 13, 607–619 (1999).
Weng, A. P. et al. Growth suppression of pre-T acute lymphoblastic leukemia cells by inhibition of notch signaling. Mol. Cell. Biol. 23, 655–664 (2003).
Miele, L., Miao, H. & Nickoloff, B. J. NOTCH signaling as a novel cancer therapeutic target. Curr. Cancer Drug Targets 6, 313–323 (2006).
Hasson, P. et al. EGFR signaling attenuates Groucho-dependent repression to antagonize Notch transcriptional output. Nature Genet. 37, 101–105 (2005).
Mizutani, K., Yoon, K., Dang, L., Tokunaga, A. & Gaiano, N. Differential Notch signalling distinguishes neural stem cells from intermediate progenitors. Nature 449, 351–355 (2007).
Schad K. et al. Continuous MEK inhibition by AZD6244 (ARRY-142886) results in exhaustion of the cutaneous keratinocytic stem cell pool and resembles senescence driven skin aging. J. Clin. Oncol. 26, (May 20 suppl.) abstr 9075 (2008).
Sibilia, M. et al. The EGF receptor provides an essential survival signal for SOS-dependent skin tumor development. Cell 102, 211–220 (2000).
Zenz, R. et al. c-Jun regulates eyelid closure and skin tumor development through EGFR signaling. Dev. Cell 4, 879–889 (2003).
Hodge, D. R. et al. Interleukin 6 supports the maintenance of p53 tumor suppressor gene promoter methylation. Cancer Res. 65, 4673–4682 (2005).
Kang, J. H. et al. Methylation in the p53 promoter is a supplementary route to breast carcinogenesis: correlation between CpG methylation in the p53 promoter and the mutation of the p53 gene in the progression from ductal carcinoma in situ to invasive ductal carcinoma. Lab. Invest. 81, 573–579 (2001).
Raman, V. et al. Compromised HOXA5 function can limit p53 expression in human breast tumours. Nature 405, 974–978 (2000).
Stuart, E. T., Haffner, R., Oren, M. & Gruss, P. Loss of p53 function through PAX-mediated transcriptional repression. EMBO J. 14, 5638–5645 (1995).
Cragg, M. S., Kuroda, J., Puthalakath, H., Huang, D. C. & Strasser, A. Gefitinib-induced killing of NSCLC cell lines expressing mutant EGFR requires BIM and can be enhanced by BH3 mimetics. PLoS Med. 4, 1681–1689 (2007).
Dotto, G. P. Signal transduction pathways controlling the switch between keratinocyte growth and differentiation. Crit. Rev. Oral Biol. Med. 10, 442–457 (1999).
Doroquez, D. B. & Rebay, I. Signal integration during development: mechanisms of EGFR and Notch pathway function and cross-talk. Crit. Rev. Biochem. Mol. Biol. 41, 339–385 (2006).
Tergaonkar, V. & Perkins, N. D. p53 and NF-κB crosstalk: IKKα tips the balance. Mol. Cell 26, 158–159 (2007).
Nguyen, B. C. et al. Cross-regulation between Notch and p63 in keratinocyte commitment to differentiation. Genes Dev. 20, 1028–1042 (2006).
Dajee, M. et al. NF-κB blockade and oncogenic Ras trigger invasive human epidermal neoplasia. Nature 421, 639–643 (2003).
Eferl, R. & Wagner, E. F. AP-1: a double-edged sword in tumorigenesis. Nature Rev. Cancer 3, 859–868 (2003).
Reuter, C. W., Morgan, M. A. & Eckardt, A. Targeting EGF-receptor-signalling in squamous cell carcinomas of the head and neck. Br. J. Cancer 96, 408–416 (2007).
Dull, T. et al. A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72, 8463–8471 (1998).
Musti, A. M., Treier, M. & Bohmann, D. Reduced ubiquitin-dependent degradation of c-Jun after phosphorylation by MAP kinases. Science 275, 400–402 (1997).
Luetteke, N. C. et al. The mouse waved-2 phenotype results from a point mutation in the EGF receptor tyrosine kinase. Genes Dev. 8, 399–413 (1994).
Acknowledgements
We thank J. Follen and C. Shamu of the Harvard Institute for Chemistry and Cell Biology, and N. Tolliday from the Broad Institute (Harvard/MIT, Cambridge, MA) for assistance with screening; W. Austen (Massachusetts General Hospital, Boston, MA) and W. Raffoul (University of Lausanne) for human skin material; N. Gaiano (John Hopkins University, Baltimore, MD) for the Notch reporter GFP mice (TNR); R. Zenz for mouse skin tumour samples; Vikram Rajashakera for technical help, and C. Brisken and C. Missero for careful reading of the manuscript. This work was supported by NIH Grants AR39190, AR 054856, the Swiss National Foundation, a grant from the European Union (Epistem, Sixth Framework Program, LSHB-CT-2005-019067) and, in part, by the Cutaneous Biology Research Center through the Massachusetts General Hospital/Shiseido Co. Agreement.
Author information
Authors and Affiliations
Contributions
V.K., A.M. and G.P.D. designed the experiments and wrote the manuscript; V.K., A.M., J.G.-V., B.H., K.L., C.L., performed the experiments; V.N., R.D. and E.F.W. provided unique sample material and conceptual insights.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary Figures S1, S2, S3, S4, S5, S6, S7 and Supplementary Tables S1, S2 and S3 (PDF 1110 kb)
Rights and permissions
About this article
Cite this article
Kolev, V., Mandinova, A., Guinea-Viniegra, J. et al. EGFR signalling as a negative regulator of Notch1 gene transcription and function in proliferating keratinocytes and cancer. Nat Cell Biol 10, 902–911 (2008). https://doi.org/10.1038/ncb1750
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1750
This article is cited by
-
Human skin specific long noncoding RNA HOXC13-AS regulates epidermal differentiation by interfering with Golgi-ER retrograde transport
Cell Death & Differentiation (2023)
-
Notch-ing up knowledge on molecular mechanisms of skin fibrosis: focus on the multifaceted Notch signalling pathway
Journal of Biomedical Science (2021)
-
Deubiquitylation and stabilization of Notch1 intracellular domain by ubiquitin-specific protease 8 enhance tumorigenesis in breast cancer
Cell Death & Differentiation (2020)
-
Immuno-detection by sequencing enables large-scale high-dimensional phenotyping in cells
Nature Communications (2018)
-
TRAF6 regulates EGF-induced cell transformation and cSCC malignant phenotype through CD147/EGFR
Oncogenesis (2018)