Treatment of BRAF(V600E) mutant melanoma by small molecule drugs that target the BRAF or MEK kinases can be effective, but resistance develops invariably1,2. In contrast, colon cancers that harbour the same BRAF(V600E) mutation are intrinsically resistant to BRAF inhibitors, due to feedback activation of the epidermal growth factor receptor (EGFR)3,4. Here we show that 6 out of 16 melanoma tumours analysed acquired EGFR expression after the development of resistance to BRAF or MEK inhibitors. Using a chromatin-regulator-focused short hairpin RNA (shRNA) library, we find that suppression of sex determining region Y-box 10 (SOX10) in melanoma causes activation of TGF-β signalling, thus leading to upregulation of EGFR and platelet-derived growth factor receptor-β (PDGFRB), which confer resistance to BRAF and MEK inhibitors. Expression of EGFR in melanoma or treatment with TGF-β results in a slow-growth phenotype with cells displaying hallmarks of oncogene-induced senescence. However, EGFR expression or exposure to TGF-β becomes beneficial for proliferation in the presence of BRAF or MEK inhibitors. In a heterogeneous population of melanoma cells having varying levels of SOX10 suppression, cells with low SOX10 and consequently high EGFR expression are rapidly enriched in the presence of drug, but this is reversed when the drug treatment is discontinued. We find evidence for SOX10 loss and/or activation of TGF-β signalling in 4 of the 6 EGFR-positive drug-resistant melanoma patient samples. Our findings provide a rationale for why some BRAF or MEK inhibitor-resistant melanoma patients may regain sensitivity to these drugs after a ‘drug holiday’ and identify patients with EGFR-positive melanoma as a group that may benefit from re-treatment after a drug holiday.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


Primary accessions

Gene Expression Omnibus

Data deposits

RNA sequencing data are available at Gene Expression Omnibus with accession code GSE50535.


  1. 1.

    et al. Improved survival with vemurafenib in melanoma with BRAF V600E mutation. N. Engl. J. Med. 364, 2507–2516 (2011)

  2. 2.

    et al. Improved survival with MEK inhibition in BRAF-mutated melanoma. N. Engl. J. Med. 367, 107–114 (2012)

  3. 3.

    et al. Unresponsiveness of colon cancer to BRAF(V600E) inhibition through feedback activation of EGFR. Nature 483, 100–103 (2012)

  4. 4.

    et al. EGFR-mediated re-activation of MAPK signaling contributes to insensitivity of BRAF mutant colorectal cancers to RAF inhibition with vemurafenib. Cancer Discov. 2, 227–235 (2012)

  5. 5.

    et al. Mutations of the BRAF gene in human cancer. Nature 417, 949–954 (2002)

  6. 6.

    , & From genes to drugs: targeted strategies for melanoma. Nature Rev. Cancer 12, 349–361 (2012)

  7. 7.

    et al. RAF inhibitor resistance is mediated by dimerization of aberrantly spliced BRAF(V600E). Nature 480, 387–390 (2011)

  8. 8.

    et al. COT drives resistance to RAF inhibition through MAP kinase pathway reactivation. Nature 468, 968–972 (2010)

  9. 9.

    et al. Dissecting therapeutic resistance to RAF inhibition in melanoma by tumour genomic profiling. J. Clin. Oncol. 29, 3085–3096 (2011)

  10. 10.

    et al. Melanoma whole-exome sequencing identifies (V600E)B-RAF amplification-mediated acquired B-RAF inhibitor resistance. Nature Commun. 3, 724 (2012)

  11. 11.

    et al. Melanomas acquire resistance to B-RAF(V600E) inhibition by RTK or N-RAS upregulation. Nature 468, 973–977 (2010)

  12. 12.

    et al. Inhibiting EGF receptor or SRC family kinase signaling overcomes BRAF inhibitor resistance in melanoma. Cancer Discov 3, 158–167 (2013)

  13. 13.

    et al. Acquired resistance to BRAF inhibitors mediated by a RAF kinase switch in melanoma can be overcome by cotargeting MEK and IGF-1R/PI3K. Cancer Cell 18, 683–695 (2010)

  14. 14.

    et al. Expression of epidermal growth factor receptor in human cultured cells and tissues: relationship to cell lineage and stage of differentiation. Cancer Res. 46, 4726–4731 (1986)

  15. 15.

    , , , & Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602 (1997)

  16. 16.

    et al. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724 (2005)

  17. 17.

    & New tools for functional mammalian cancer genetics. Nature Rev. Cancer 3, 781–789 (2003)

  18. 18.

    et al. MED12 controls the response to multiple cancer drugs through regulation of TGF-β receptor signaling. Cell 151, 937–950 (2012)

  19. 19.

    et al. Activator protein-1 mediates induced but not basal epidermal growth factor receptor gene expression. Mol. Med. 6, 17–27 (2000)

  20. 20.

    et al. c-Jun regulates eyelid closure and skin tumor development through EGFR signaling. Dev. Cell 4, 879–889 (2003)

  21. 21.

    et al. PDGFRB promotes liver metastasis formation of mesenchymal-like colorectal tumor cells. Neoplasia 15, 204–217 (2013)

  22. 22.

    , & Smad3 and Smad4 cooperate with c-Jun/c-Fos to mediate TGF-β-induced transcription. Nature 394, 909–913 (1998)

  23. 23.

    et al. DNA topoisomerase I is a cofactor for c-Jun in the regulation of epidermal growth factor receptor expression and cancer cell proliferation. Mol. Cell. Biol. 25, 5040–5051 (2005)

  24. 24.

    et al. Interaction among SOX10, PAX3 and MITF, three genes altered in Waardenburg syndrome. Hum. Mol. Genet. 9, 1907–1917 (2000)

  25. 25.

    et al. Systematic identification of genomic markers of drug sensitivity in cancer cells. Nature 483, 570–575 (2012)

  26. 26.

    , , & Successful rechallenge in two patients with BRAF-V600-mutant melanoma who experienced previous progression during treatment with a selective BRAF inhibitor. Melanoma Res. 22, 466–472 (2012)

  27. 27.

    et al. A public genome-scale lentiviral expression library of human ORFs. Nature Methods 8, 659–661 (2011)

  28. 28.

    et al. Frequent mutations in the MITF pathway in melanoma. Pigment Cell Melanoma Res 22, 435–444 (2009)

  29. 29.

    et al. ZNF423 is critically required for retinoic acid-induced differentiation and is a marker of neuroblastoma outcome. Cancer Cell 15, 328–340 (2009)

Download references


We thank the NKI Core Facilities for Genomics and Molecular Pathology & Biobanking for tumour tissue and support in DNA sequencing. We thank S. Roy for collecting clinical data and N. Kamsu Kom for tissue preparation. This work was supported by grants from the European Research Council (ERC), the Dutch Cancer Society (KWF), the EU COLTHERES project and grants by the Netherlands Organization for Scientific Research (NWO) to Cancer Genomics Netherlands (CGC.NL). Additional support was provided by Fondazione Piemontese per la Ricerca sul Cancro—ONLUS grant ‘Farmacogenomica—5 per mille 2009 MIUR’ (F.D.N.); AIRC MFAG 11349 (F.D.N.); AIRC IG grant n. 12812 (A.B.); and Canadian Institutes of Health Research (CIHR) grant MOP-130540 (S.Hu).

Author information

Author notes

    • Chong Sun
    • , Liqin Wang
    •  & Sidong Huang

    These authors contributed equally to this work.


  1. Division of Molecular Carcinogenesis, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands

    • Chong Sun
    • , Liqin Wang
    • , Sidong Huang
    • , Guus J. J. E. Heynen
    • , Anirudh Prahallad
    • , Stefan M. Willems
    • , Prashanth K. Bajpe
    • , Cor Lieftink
    • , Wipawadee Grernrum
    • , Andreas Schlicker
    • , Lodewyk F. A. Wessels
    • , Roderick L. Beijersbergen
    •  & Rene Bernards
  2. Department of Biochemistry, The Rosalind and Morris Goodman Cancer Centre, McGill University, Montreal, Quebec H3G 1Y6, Canada

    • Sidong Huang
  3. Institut Gustave Roussy, 114 Rue Edouard Vaillant, 94800 Villejuif, France

    • Caroline Robert
    • , Christina Mateus
    • , Stephan Vagner
    •  & Alexander M. M. Eggermont
  4. Division of Medical Oncology, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands

    • John Haanen
    •  & Christian Blank
  5. Division of Pathology, Cancer Systems Biology Centre and Cancer Genomics Centre Netherlands, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands

    • Jelle Wesseling
    •  & Ingrid Hofland
  6. Department of Pathology, University Medical Centre Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands

    • Stefan M. Willems
  7. University of Torino, Department of Oncology, Str prov 142 Km 3.95, 10060 Candiolo, Torino, Italy

    • Davide Zecchin
    • , Alberto Bardelli
    •  & Federica Di Nicolantonio
  8. Candiolo Cancer Institute – FPO, IRCCS, Str prov 142 Km 3.95, 10060 Candiolo, Torino, Italy

    • Davide Zecchin
    • , Sebastijan Hobor
    • , Alberto Bardelli
    •  & Federica Di Nicolantonio
  9. FIRC Institute of Molecular Oncology (IFOM), 20139 Milano, Italy

    • Alberto Bardelli


  1. Search for Chong Sun in:

  2. Search for Liqin Wang in:

  3. Search for Sidong Huang in:

  4. Search for Guus J. J. E. Heynen in:

  5. Search for Anirudh Prahallad in:

  6. Search for Caroline Robert in:

  7. Search for John Haanen in:

  8. Search for Christian Blank in:

  9. Search for Jelle Wesseling in:

  10. Search for Stefan M. Willems in:

  11. Search for Davide Zecchin in:

  12. Search for Sebastijan Hobor in:

  13. Search for Prashanth K. Bajpe in:

  14. Search for Cor Lieftink in:

  15. Search for Christina Mateus in:

  16. Search for Stephan Vagner in:

  17. Search for Wipawadee Grernrum in:

  18. Search for Ingrid Hofland in:

  19. Search for Andreas Schlicker in:

  20. Search for Lodewyk F. A. Wessels in:

  21. Search for Roderick L. Beijersbergen in:

  22. Search for Alberto Bardelli in:

  23. Search for Federica Di Nicolantonio in:

  24. Search for Alexander M. M. Eggermont in:

  25. Search for Rene Bernards in:


R.B., A.B., F.D.N., L.F.A.W., C.R., R.L.B. and A.M.M.E. supervised all research. R.B. and C.S. wrote the manuscript. C.S., L.W., S.Hu., G.J.J.E.H., A.P., D.Z., S.Ho., P.K.B., C.L., C.M., S.V., J.W., W.G., I.H. and A.S. designed and performed experiments and J.H., C.B., C.R., S.M.W., S.V. and A.M.M.E. provided clinical samples and gave advice.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Rene Bernards.

Extended data

Supplementary information

Excel files

  1. 1.

    Supplementary Table 1

    This table contains patient information.

  2. 2.

    Supplementary Table 2

    A list of genes in chromatin library.

  3. 3.

    Supplementary Table 3

    Top hits from genetic screen.

  4. 4.

    Supplementary Table 4

    RNAseq data from shSOX10 cells.

  5. 5.

    Supplementary Table 5

    Gene Set Enrichment Analysis of SOX10 regulated genes.

  6. 6.

    Supplementary Table 6

    shRNA IDs and sequences.

  7. 7.

    Supplementary Table 7

    List of QRT-PCR primers used.

About this article

Publication history





Further reading


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.