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:

Early epigenetic downregulation of WNK2 kinase during pancreatic ductal adenocarcinoma development

Oncogene volumeĀ 33,Ā pages 3401ā€“3410 (2014)Cite this article

Subjects

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is usually incurable. Contrary to genetic mechanisms involved in PDAC pathogenesis, epigenetic alterations are ill defined. Here, we determine the contribution of epigenetically silenced genes to the development of PDAC. We analyzed enriched, highly methylated DNAs from PDACs, chronic pancreatitis (CP) and normal tissues using CpG island microarrays and identified WNK2 as a prominent candidate tumor suppressor gene being downregulated early in PDAC development. WNK2 was further investigated in tissue microarrays, methylation analysis of early pancreatic intraepithelial neoplasia (PanIN), mouse models for PDAC and pancreatitis, re-expression studies after demethylation, and cell growth assays using WNK2 overexpression. Demethylation assays confirmed the link between methylation and expression. WNK2 hypermethylation was higher in tumor than in surrounding inflamed tissues and was observed in PanIN lesions as well as in a PDAC mouse model. WNK2 mRNA and protein expressions were lower in PDAC and CP compared with normal tissues both in patients and mouse models. Overexpression of WNK2 led to reduced cell growth, and WNK2 expression in tissues correlated negatively with pERK1/2 expression, a downstream target of WNK2 responsible for cell proliferation. Downregulation of WNK2 by promoter hypermethylation occurs early in PDAC pathogenesis and may support tumor cell growth via the ERKā€“MAPK pathway.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Ghaneh P, Costello E, Neoptolemos JP . Biology and management of pancreatic cancer. Postgrad Med J 2008; 84: 478ā€“497.

    ArticleĀ  CASĀ  Google ScholarĀ 

  2. Bardeesy N, DePinho RA . Pancreatic cancer biology and genetics. Nat Rev Cancer 2002; 2: 897ā€“909.

    ArticleĀ  CASĀ  Google ScholarĀ 

  3. Hezel AF, Kimmelman AC, Stanger BZ, Bardeesy N, DePinho RA . Genetics and biology of pancreatic ductal adenocarcinoma. Genes Dev 2006; 20: 1218ā€“1249.

    ArticleĀ  CASĀ  Google ScholarĀ 

  4. Goggins M . Molecular markers of early pancreatic cancer. J Clin Oncol 2005; 23: 4524ā€“4531.

    ArticleĀ  CASĀ  Google ScholarĀ 

  5. Sato N, Fukushima N, Maitra A, Matsubayashi H, Yeo CJ, Cameron JL et al. Discovery of novel targets for aberrant methylation in pancreatic carcinoma using high-throughput microarrays. Cancer Res 2003; 63: 3735ā€“3742.

    CASĀ  Google ScholarĀ 

  6. Omura N, Li CP, Li A, Hong SM, Walter K, Jimeno A et al. Genome-wide profiling of methylated promoters in pancreatic adenocarcinoma. Cancer Biol Ther 2008; 7: 1146ā€“1156.

    ArticleĀ  CASĀ  Google ScholarĀ 

  7. Vincent A, Omura N, Hong S-M, Jaffe A, Eshleman J, Goggins M . Genome-wide analysis of promoter methylation associated with gene expression profile in pancreatic adenocarcinoma. Clin Cancer Res 2011; 17: 4341ā€“4354.

    ArticleĀ  CASĀ  Google ScholarĀ 

  8. Sipos B, Frank S, Gress T, Hahn S, Klƶppel G . Pancreatic intraepithelial neoplasia revisited and updated. Pancreatology 2009; 9: 45ā€“54.

    ArticleĀ  CASĀ  Google ScholarĀ 

  9. Buchholz M, Braun M, Heidenblut A, Kestler HA, Klƶppel G, Schmiegel W et al. Transcriptome analysis of microdissected pancreatic intraepithelial neoplastic lesions. Oncogene 2005; 24: 6626ā€“6636.

    ArticleĀ  CASĀ  Google ScholarĀ 

  10. Sato N, Fukushima N, Hruban RH, Goggins M . CpG island methylation profile of pancreatic intraepithelial neoplasia. Mod Pathol 2008; 21: 238ā€“244.

    ArticleĀ  CASĀ  Google ScholarĀ 

  11. Jansen M, Fukushima N, Rosty C, Walter K, Altink R, Heek Tv et al. Aberrant methylation of the 5' CpG island of TSLC1 is common in pancreatic ductal adenocarcinoma and is first manifest in high-grade PanINs. Cancer Biol Ther 2002; 1: 284ā€“287.

    ArticleĀ  Google ScholarĀ 

  12. Fukushima N, Sato N, Ueki T, Rosty C, Walter KM, Wilentz RE et al. Aberrant Methylation of Preproenkephalin and p16 Genes in Pancreatic Intraepithelial Neoplasia and Pancreatic Ductal Adenocarcinoma. Am J Pathol 2002; 160: 1573ā€“1581.

    ArticleĀ  CASĀ  Google ScholarĀ 

  13. Peng D-F, Kanai Y, Sawada M, Ushijima S, Hiraoka N, Kitazawa S et al. DNA methylation of multiple tumor-related genes in association with overexpression of DNA methyltransferase 1 (DNMT1) during multistage carcinogenesis of the pancreas. Carcinogenesis 2006; 27: 1160ā€“1168.

    ArticleĀ  CASĀ  Google ScholarĀ 

  14. Lowenfels AB, Maisonneuve P, Cavallini G, Ammann RW, Lankisch PG, Andersen JR et al. Pancreatitis and the Risk of Pancreatic Cancer. N Engl J Med 1993; 328: 1433ā€“1437.

    ArticleĀ  CASĀ  Google ScholarĀ 

  15. Matsubayashi H, Canto M, Sato N, Klein A, Abe T, Yamashita K et al. DNA methylation alterations in the pancreatic juice of patients with suspected pancreatic disease. Cancer Res 2006; 66: 1208ā€“1217.

    ArticleĀ  CASĀ  Google ScholarĀ 

  16. Gold DV, Goggins M, Modrak DE, Newsome G, Liu M, Shi C et al. Detection of Early-Stage Pancreatic Adenocarcinoma. Cancer Epidemiol Biomarkers Prev 2010; 19: 2786ā€“2794.

    ArticleĀ  CASĀ  Google ScholarĀ 

  17. Liggett T, Melnikov A, Yi Q-l, Replogle C, Brand R, Kaul K et al. Differential methylation of cell-free circulating DNA among patients with pancreatic cancer versus chronic pancreatitis. Cancer 2010; 116: 1674ā€“1680.

    ArticleĀ  CASĀ  Google ScholarĀ 

  18. Schilling E, Rehli M . Global, comparative analysis of tissue-specific promoter CpG methylation. Genomics 2007; 90: 314ā€“323.

    ArticleĀ  CASĀ  Google ScholarĀ 

  19. Moniz S, Matos P, Jordan P . WNK2 modulates MEK1 activity through the Rho GTPase pathway. Cellular Signalling 2008; 20: 1762ā€“1768.

    ArticleĀ  CASĀ  Google ScholarĀ 

  20. Moniz S, VerĆ­ssimo F, Matos P, BrazĆ£o R, Silva E, Kotevelets L et al. Protein kinase WNK2 inhibits cell proliferation by negatively modulating the activation of MEK1/ERK1/2. Oncogene 2007; 26: 6071ā€“6081.

    ArticleĀ  CASĀ  Google ScholarĀ 

  21. Hong C, Moorefield KS, Jun P, Aldape KD, Kharbanda S, Phillips HS et al. Epigenome scans and cancer genome sequencing converge on WNK2, a kinase-independent suppressor of cell growth. Proc Natl Acad Sci USA 2007; 104: 10974ā€“10979.

    ArticleĀ  CASĀ  Google ScholarĀ 

  22. Jun P, Hong C, Lal A, Wong JM, McDermott MW, Bollen AW et al. Epigenetic silencing of the kinase tumor suppressor WNK2 is tumor-type and tumor-grade specific. Neuro-Oncology 2008; 11: 414ā€“422.

    ArticleĀ  Google ScholarĀ 

  23. Lenertz LY, Lee B-H, Min X, Xu B-e, Wedin K, Earnest S et al. Properties of WNK1 and implications for other family members. J Biol Chem 2005; 280: 26653ā€“26658.

    ArticleĀ  CASĀ  Google ScholarĀ 

  24. Xu B-e, Stippec S, Lenertz L, Lee B-H, Zhang W, Lee Y-K et al. WNK1 activates ERK5 by an MEKK2/3-dependent mechanism. J Biol Chem 2004; 279: 7826ā€“7831.

    ArticleĀ  CASĀ  Google ScholarĀ 

  25. Sun X, Gao L, Yu RK, Zeng G . Down-regulation of WNK1 protein kinase in neural progenitor cells suppresses cell proliferation and migration. J Neurochem 2006; 99: 1114ā€“1121.

    ArticleĀ  CASĀ  Google ScholarĀ 

  26. Haas BR, Cuddapah VA, Watkins S, Rohn KJ, Dy TE, Sontheimer H . With-No-Lysine Kinase 3 (WNK3) stimulates glioma invasion by regulating cell volume. Am J Physiologyā€”Cell Physiol 2011; 301: C1150ā€“C1160.

    ArticleĀ  CASĀ  Google ScholarĀ 

  27. Grapin-Botton A . Ductal cells of the pancreas. Int J Biochem Cell Biol 2005; 37: 504ā€“510.

    ArticleĀ  CASĀ  Google ScholarĀ 

  28. Heller A, Zƶrnig I, MĆ¼ller T, Giorgadze K, Frei C, Giese T et al. Immunogenicity of SEREX-identified antigens and disease outcome in pancreatic cancer. Cancer Immunol, Immunother 2010; 59: 1389ā€“1400.

    ArticleĀ  CASĀ  Google ScholarĀ 

  29. Kong B, Michalski CW, Erkan M, Friess H, Kleeff J . From tissue turnover to the cell of origin for pancreatic cancer. Nat Rev Gastroenterol Hepatol 2011; 8: 467ā€“472.

    ArticleĀ  Google ScholarĀ 

  30. Meloche S, Pouyssegur J . The ERK1//2 mitogen-activated protein kinase pathway as a master regulator of the G1- to S-phase transition. Oncogene 2007; 26: 3227ā€“3239.

    ArticleĀ  CASĀ  Google ScholarĀ 

  31. Hartwig W, Hackert T, Hinz U, Gluth A, Bergmann F, Strobel O et al. Pancreatic cancer surgery in the new millennium: better prediction of outcome. Ann Surg 2011; 254: 311ā€“319.

    ArticleĀ  Google ScholarĀ 

  32. Strobel O, Dor Y, Stirman A, Trainor A, FernĆ”ndez-del Castillo C, Warshaw AL et al. Beta cell transdifferentiation does not contribute to preneoplastic/metaplastic ductal lesions of the pancreas by genetic lineage tracing in vivo. Proc Natl Acad Sci 2007; 104: 4419ā€“4424.

    ArticleĀ  CASĀ  Google ScholarĀ 

  33. Niederau C, Ferrell L, Grendell J . Caerulein-induced acute necrotizing pancreatitis in mice: protective effects of proglumide, benzotript, and secretin. Gastroenterology 1985; 88: 1192ā€“1204.

    ArticleĀ  CASĀ  Google ScholarĀ 

  34. Jensen JN, Cameron E, Garay MVR, Starkey TW, Gianani R, Jensen J . Recapitulation of elements of embryonic development in adult mouse pancreatic regeneration. Gastroenterology 2005; 128: 728ā€“741.

    ArticleĀ  CASĀ  Google ScholarĀ 

  35. Pinho AV, Rooman I, Reichert M, De Medts N, Bouwens L, Rustgi AK et al. Adult pancreatic acinar cells dedifferentiate to an embryonic progenitor phenotype with concomitant activation of a senescence programme that is present in chronic pancreatitis. Gut 2011; 60: 958ā€“966.

    ArticleĀ  CASĀ  Google ScholarĀ 

  36. Means AL, Meszoely IM, Suzuki K, Miyamoto Y, Rustgi AK, Coffey RJ et al. Pancreatic epithelial plasticity mediated by acinar cell transdifferentiation and generation of nestin-positive intermediates. Development 2005; 132: 3767ā€“3776.

    ArticleĀ  CASĀ  Google ScholarĀ 

  37. Hingorani SR, Petricoin Iii EF, Maitra A, Rajapakse V, King C, Jacobetz MA et al. Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell 2003; 4: 437ā€“450.

    ArticleĀ  CASĀ  Google ScholarĀ 

  38. Kuhmann C, Weichenhan D, Rehli M, Plass C, Schmezer P, Popanda O . DNA methylation changes in cells regrowing after fractioned ionizing radiation. Radiother Oncol 2011; 101: 116ā€“121.

    ArticleĀ  CASĀ  Google ScholarĀ 

  39. Ehrich M, Nelson MR, Stanssens P, Zabeau M, Liloglou T, Xinarianos G et al. Quantitative high-throughput analysis of DNA methylation patterns by base-specific cleavage and mass spectrometry. PNAS 2005; 102: 15785ā€“15790.

    ArticleĀ  CASĀ  Google ScholarĀ 

Download references

Acknowledgements

We would like to thank Reinhard Gliniorz, Oliver MĆ¼cke, Jana Petersen, Peter Waas and Otto Zelezny for their technical support, Dr Marta Faryna and Dr Carolin Konermann for discussions regarding the MCIp/microarray assays, Anna Poetsch and Christopher Schmidt for experimental advices, Dr Rainer Claus and Dr Christopher Oakes for stimulating discussions and Dr Ming-Sound Tsao (Ontario Cancer Institute, Toronto, Canada) for providing us the HPDE6-E6E7 cell line. This work is supported by grants from the Cancer Institute New South Wales, Australia (10FRL203 fellowship), from the German Federal Ministry of Education and Research (PKB-01GS08114) and from the German Federal Ministry of Education and Research as part of the PaCaNet consortium within the NGFN program. We thank the Fulbright commission in Germany and the US State Department for providing external funding (TK).

Author information

Authors and Affiliations

  1. Division of Epigenomics and Cancer Risk Factors, German Cancer Research Center (DKFZ), Heidelberg, Germany

    C Dutruel,Ā D Weichenhan,Ā L Geiselhart,Ā T Kaffenberger,Ā C Plass,Ā O PopandaĀ &Ā P Schmezer

  2. Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany

    F BergmannĀ &Ā P Schirmacher

  3. Cancer Research Program, Garvan Institute of Medical Research, Darlinghurst, Australia

    I Rooman

  4. Division of Biostatistics, German Cancer Research Center (DKFZ), Heidelberg, Germany

    M Zucknick

  5. Division of Molecular Genetic Epidemiology, German Cancer Research Center (DKFZ), Heidelberg, Germany

    P S Rachakonda,Ā H XieĀ &Ā R Kumar

  6. Functional Genome Analysis, German Cancer Research Center (DKFZ), Heidelberg, Germany

    A BauerĀ &Ā J Hoheisel

  7. Department of General, Visceral and Transplantation Surgery, University of Heidelberg, Heidelberg, Germany

    N GieseĀ &Ā J Werner

  8. Department of Neurosurgery, Brain Tumor Research Center, Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, California, USA

    C HongĀ &Ā J F Costello

  9. Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany

    M Rehli

Authors
  1. C Dutruel
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  2. F Bergmann
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  3. I Rooman
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  4. M Zucknick
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  5. D Weichenhan
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  6. L Geiselhart
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  7. T Kaffenberger
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  8. P S Rachakonda
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  9. A Bauer
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  10. N Giese
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  11. C Hong
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  12. H Xie
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  13. J F Costello
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  14. J Hoheisel
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  15. R Kumar
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  16. M Rehli
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  17. P Schirmacher
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  18. J Werner
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  19. C Plass
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  20. O Popanda
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

  21. P Schmezer
    View author publications

    You can also search for this author in PubMedĀ Google Scholar

Corresponding author

Correspondence to P Schmezer.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dutruel, C., Bergmann, F., Rooman, I. et al. Early epigenetic downregulation of WNK2 kinase during pancreatic ductal adenocarcinoma development. Oncogene 33, 3401ā€“3410 (2014). https://doi.org/10.1038/onc.2013.312

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2013.312

Keywords

This article is cited by

Search

Advanced search

Quick links