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:

TGFβ promotes mesenchymal phenotype of pancreatic cancer cells, in part, through epigenetic activation of VAV1

Oncogene volume 36, pages 2202–2214 (2017)Cite this article

Subjects

Abstract

The highly homeostasis-resistant nature of cancer cells leads to their escape from treatment and to liver metastasis, which in turn makes pancreatic ductal adenocarcinoma (PDAC) difficult to treat, especially the squamous/epithelial-to-mesenchymal transition (EMT)-like subtype. As the molecular mechanisms underlying tumour heterogeneity remain elusive, we investigated whether epigenetic regulation might explain inter-individual differences in the progression of specific subtypes. DNA methylation profiling performed on cancer tissues prior to chemo/radiotherapy identified one hypermethylated CpG site (CpG6882469) in the VAV1 gene body that was correlated with demethylation of two promoter CpGs (CpG6772370/CpG6772811) in both PDAC and peripheral blood. Transforming growth factor β treatment induced gene-body hypermethylation, dissociation of DNMT1 from the promoter, and VAV1 expression via SMAD4 and mutant KrasG12D. Pharmacological inhibition of TGFβ-VAV1 signalling decreased the squamous/EMT-like cancer cells, promoted nuclear VAV1 localization, and enhanced the efficacy of gemcitabine in prolonging the survival of KPfl/flC mice. Together, the three VAV1 CpGs serve as biomarkers for prognosis and early detection, and the TGFβ-VAV1 axis represents a therapeutic target.

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
Figure 8

Similar content being viewed by others

References

  1. Achyut BR, Yang L . Transforming growth factor-beta in the gastrointestinal and hepatic tumor microenvironment. Gastroenterology 2011; 141: 1167–1178.

    Article  CAS  PubMed  Google Scholar 

  2. Collazo J, Zhu B, Larkin S, Martin SK, Pu H, Horbinski C et al. Cofilin drives cell-invasive and metastatic responses to TGF-beta in prostate cancer. Cancer Res 2014; 74: 2362–2373.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Tufegdzic Vidakovic A, Rueda OM, Vervoort SJ, Sati Batra A, Goldgraben MA, Uribe-Lewis S et al. Context-specific effects of TGF-beta/SMAD3 in cancer are modulated by the epigenome. Cell Rep 2015; 13: 2480–2490.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Hezel AF, Deshpande V, Zimmerman SM, Contino G, Alagesan B, O'Dell MR et al. TGF-beta and alphavbeta6 integrin act in a common pathway to suppress pancreatic cancer progression. Cancer Res 2012; 72: 4840–4845.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Nakajima S, Doi R, Toyoda E, Tsuji S, Wada M, Koizumi M et al. N-cadherin expression and epithelial-mesenchymal transition in pancreatic carcinoma. Clin Cancer Res 2004; 10 (12 Pt 1): 4125–4133.

    Article  CAS  PubMed  Google Scholar 

  6. Ebert MP, Fei G, Schandl L, Mawrin C, Dietzmann K, Herrera P et al. Reduced PTEN expression in the pancreas overexpressing transforming growth factor-beta 1. Br J Cancer 2002; 86: 257–262.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Gore AJ, Deitz SL, Palam LR, Craven KE, Korc M . Pancreatic cancer-associated retinoblastoma 1 dysfunction enables TGF-beta to promote proliferation. J Clin Invest 2014; 124: 338–352.

    Article  CAS  PubMed  Google Scholar 

  8. Gaspar NJ, Li L, Kapoun AM, Medicherla S, Reddy M, Li G et al. Inhibition of transforming growth factor beta signaling reduces pancreatic adenocarcinoma growth and invasiveness. Mol Pharmacol 2007; 72: 152–161.

    Article  CAS  PubMed  Google Scholar 

  9. Ostapoff KT, Cenik BK, Wang M, Ye R, Xu X, Nugent D et al. Neutralizing murine TGFbetaR2 promotes a differentiated tumor cell phenotype and inhibits pancreatic cancer metastasis. Cancer Res 2014; 74: 4996–5007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Katzav S . Vav1: A Dr. Jekyll and Mr. Hyde protein – good for the hematopoietic system, bad for cancer. Oncotarget 2015; 6: 28731–28742.

    PubMed  PubMed Central  Google Scholar 

  11. Katzav S, Martin-Zanca D, Barbacid M . vav, a novel human oncogene derived from a locus ubiquitously expressed in hematopoietic cells. EMBO J 1989; 8: 2283–2290.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Qi Y, Kong FM, Deng Q, Li JY, Cui R, Pu YD et al. Clinical significance and prognostic value of Vav1 expression in Non-small cell lung cancer. Am J Cancer Res 2015; 5: 2491–2497.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Sebban S, Farago M, Rabinovich S, Lazer G, Idelchuck Y, Ilan L et al. Vav1 promotes lung cancer growth by instigating tumor-microenvironment cross-talk via growth factor secretion. Oncotarget 2014; 5: 9214–9226.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Grassilli S, Brugnoli F, Lattanzio R, Rossi C, Perracchio L, Mottolese M et al. High nuclear level of Vav1 is a positive prognostic factor in early invasive breast tumors: a role in modulating genes related to the efficiency of metastatic process. Oncotarget 2014; 5: 4320–4336.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Lindsey JC, Kawauchi D, Schwalbe EC, Solecki DJ, Selby MP, McKinnon PJ et al. Cross-species epigenetics identifies a critical role for VAV1 in SHH subgroup medulloblastoma maintenance. Oncogene 2015; 34: 4746–4757.

    Article  CAS  PubMed  Google Scholar 

  16. Garcia JL, Couceiro J, Gomez-Moreta JA, Gonzalez Valero JM, Briz AS, Sauzeau V et al. Expression of VAV1 in the tumour microenvironment of glioblastoma multiforme. J Neurooncol 2012; 110: 69–77.

    Article  CAS  PubMed  Google Scholar 

  17. Kataoka K, Nagata Y, Kitanaka A, Shiraishi Y, Shimamura T, Yasunaga J et al. Integrated molecular analysis of adult T cell leukemia/lymphoma. Nat Genet 2015; 47: 1304–1315.

    Article  CAS  PubMed  Google Scholar 

  18. Bertagnolo V, Brugnoli F, Mischiati C, Sereni A, Bavelloni A, Carini C et al. Vav promotes differentiation of human tumoral myeloid precursors. Exp Cell Res 2005; 306: 56–63.

    Article  CAS  PubMed  Google Scholar 

  19. Brugnoli F, Lambertini E, Varin-Blank N, Piva R, Marchisio M, Grassilli S et al. Vav1 and PU.1 are recruited to the CD11b promoter in APL-derived promyelocytes: role of Vav1 in modulating PU.1-containing complexes during ATRA-induced differentiation. Exp Cell Res 2010; 316: 38–47.

    Article  CAS  PubMed  Google Scholar 

  20. Crespo P, Schuebel KE, Ostrom AA, Gutkind JS, Bustelo XR . Phosphotyrosine-dependent activation of Rac-1 GDP/GTP exchange by the vav proto-oncogene product. Nature 1997; 385: 169–172.

    Article  CAS  PubMed  Google Scholar 

  21. Margolis B, Hu P, Katzav S, Li W, Oliver JM, Ullrich A et al. Tyrosine phosphorylation of vav proto-oncogene product containing SH2 domain and transcription factor motifs. Nature 1992; 356: 71–74.

    Article  CAS  PubMed  Google Scholar 

  22. Bustelo XR, Ledbetter JA, Barbacid M . Product of vav proto-oncogene defines a new class of tyrosine protein kinase substrates. Nature 1992; 356: 68–71.

    Article  CAS  PubMed  Google Scholar 

  23. Razanadrakoto L, Cormier F, Lauriente V, Dondi E, Gardano L, Katzav S et al. Mutation of Vav1 adaptor region reveals a new oncogenic activation. Oncotarget 2015; 6: 2524–2537.

    Article  PubMed  Google Scholar 

  24. Razidlo GL, Wang Y, Chen J, Krueger EW, Billadeau DD, McNiven MA . Dynamin 2 potentiates invasive migration of pancreatic tumor cells through stabilization of the Rac1 GEF Vav1. Dev Cell 2013; 24: 573–585.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Nolz JC, Gomez TS, Billadeau DD . The Ezh2 methyltransferase complex: actin up in the cytosol. Trends Cell Biol 2005; 15: 514–517.

    Article  CAS  PubMed  Google Scholar 

  26. Su IH, Dobenecker MW, Dickinson E, Oser M, Basavaraj A, Marqueron R et al. Polycomb group protein ezh2 controls actin polymerization and cell signaling. Cell 2005; 121: 425–436.

    Article  CAS  PubMed  Google Scholar 

  27. Gunawan M, Venkatesan N, Loh JT, Wong JF, Berger H, Neo WH et al. The methyltransferase Ezh2 controls cell adhesion and migration through direct methylation of the extranuclear regulatory protein talin. Nat Immunol 2015; 16: 505–516.

    Article  CAS  PubMed  Google Scholar 

  28. Fernandez-Zapico ME, Gonzalez-Paz NC, Weiss E, Savoy DN, Molina JR, Fonseca R et al. Ectopic expression of VAV1 reveals an unexpected role in pancreatic cancer tumorigenesis. Cancer Cell 2005; 7: 39–49.

    Article  CAS  PubMed  Google Scholar 

  29. Yang X, Han H, De Carvalho DD, Lay FD, Jones PA, Liang G . Gene body methylation can alter gene expression and is a therapeutic target in cancer. Cancer Cell 2014; 26: 577–590.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Jones PA . Functions of DNA methylation: islands, start sites, gene bodies and beyond. Nat Rev Genet 2012; 13: 484–492.

    Article  CAS  PubMed  Google Scholar 

  31. Grimmer MR, Farnham PJ . Can genome engineering be used to target cancer-associated enhancers? Epigenomics 2014; 6: 493–501.

    Article  CAS  PubMed  Google Scholar 

  32. Hou YC, Chao YJ, Tung HL, Wang HC, Shan YS . Coexpression of CD44-positive/CD133-positive cancer stem cells and CD204-positive tumor-associated macrophages is a predictor of survival in pancreatic ductal adenocarcinoma. Cancer 2014; 120: 2766–2777.

    Article  CAS  PubMed  Google Scholar 

  33. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005; 102: 15545–15550.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Bardeesy N, Aguirre AJ, Chu GC, Cheng KH, Lopez LV, Hezel AF et al. Both p16(Ink4a) and the p19(Arf)-p53 pathway constrain progression of pancreatic adenocarcinoma in the mouse. Proc Natl Acad Sci USA 2006; 103: 5947–5952.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Bailey P, Chang DK, Nones K, Johns AL, Patch AM, Gingras MC et al. Genomic analyses identify molecular subtypes of pancreatic cancer. Nature 2016; 531: 47–52.

    Article  CAS  PubMed  Google Scholar 

  36. Thillainadesan G, Chitilian JM, Isovic M, Ablack JN, Mymryk JS, Tini M et al. TGF-beta-dependent active demethylation and expression of the p15ink4b tumor suppressor are impaired by the ZNF217/CoREST complex. Mol Cell 2012; 46: 636–649.

    Article  CAS  PubMed  Google Scholar 

  37. Ksionda O, Saveliev A, Kochl R, Rapley J, Faroudi M, Smith-Garvin JE et al. Mechanism and function of Vav1 localisation in TCR signalling. J Cell Sci 2012; 125 (Pt 22): 5302–5314.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Houlard M, Arudchandran R, Regnier-Ricard F, Germani A, Gisselbrecht S, Blank U et al. Vav1 is a component of transcriptionally active complexes. J Exp Med 2002; 195: 1115–1127.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Houlard M, Romero-Portillo F, Germani A, Depaux A, Regnier-Ricard F, Gisselbrecht S et al. Characterization of VIK-1: a new Vav-interacting Kruppel-like protein. Oncogene 2005; 24: 28–38.

    Article  CAS  PubMed  Google Scholar 

  40. Dong Z, Davidson D, Perez-Quintero LA, Kurosaki T, Swat W, Veillette A . The adaptor SAP controls NK cell activation by regulating the enzymes Vav-1 and SHIP-1 and by enhancing conjugates with target cells. Immunity 2012; 36: 974–985.

    Article  CAS  PubMed  Google Scholar 

  41. Reynolds LF, Smyth LA, Norton T, Freshney N, Downward J, Kioussis D et al. Vav1 transduces T cell receptor signals to the activation of phospholipase C-gamma1 via phosphoinositide 3-kinase-dependent and -independent pathways. J Exp Med 2002; 195: 1103–1114.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Miletic AV, Graham DB, Sakata-Sogawa K, Hiroshima M, Hamann MJ, Cemerski S et al. Vav links the T cell antigen receptor to the actin cytoskeleton and T cell activation independently of intrinsic guanine nucleotide exchange activity. PLoS One 2009; 4: e6599.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Saveliev A, Vanes L, Ksionda O, Rapley J, Smerdon SJ, Rittinger K et al. Function of the nucleotide exchange activity of vav1 in T cell development and activation. Sci Signal 2009; 2: ra83.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Razidlo GL, Magnine C, Sletten AC, Hurley RM, Almada LL, Fernandez-Zapico ME et al. Targeting pancreatic cancer metastasis by inhibition of Vav1, a driver of tumor cell invasion. Cancer Res 2015; 75: 2907–2915.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Li R, Wei F, Yu J, Li H, Ren X, Hao X . IDO inhibits T-cell function through suppressing Vav1 expression and activation. Cancer Bio Ther 2009; 8: 1402–1408.

    Article  CAS  Google Scholar 

  46. Tsai JH, Yang J . Epithelial-mesenchymal plasticity in carcinoma metastasis. Genes Dev 2013; 27: 2192–2206.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. 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. Proc Natl Acad Sci USA 2005; 102: 15785–15790.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Krohn A, Diedler T, Burkhardt L, Mayer PS, De Silva C, Meyer-Kornblum M et al. Genomic deletion of PTEN is associated with tumor progression and early PSA recurrence in ERG fusion-positive and fusion-negative prostate cancer. Am J Pathol 2012; 181: 401–412.

    Article  CAS  PubMed  Google Scholar 

  49. Huang PH, Chen CH, Chou CC, Sargeant AM, Kulp SK, Teng CM et al. Histone deacetylase inhibitors stimulate histone H3 lysine 4 methylation in part via transcriptional repression of histone H3 lysine 4 demethylases. Mol Pharmacol 2011; 79: 197–206.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Huang PH, Chuang HC, Chou CC, Wang H, Lee SL, Yang HC et al. Vitamin E facilitates the inactivation of the kinase Akt by the phosphatase PHLPP1. Sci Signal 2013; 6: ra19.

    PubMed  PubMed Central  Google Scholar 

  51. Bai LY, Weng JR, Chiu CF, Wu CY, Yeh SP, Sargeant AM et al. OSU-A9, an indole-3-carbinol derivative, induces cytotoxicity in acute myeloid leukemia through reactive oxygen species-mediated apoptosis. Biochem Pharmacol 2013; 86: 1430–1440.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors acknowledge the assistance of Yi-Chen Chen, Pei-Rong Gu, and Hui-Ling Tung, as well as assistance from the following core facilities: the Clinical Medicine Research Center of National Cheng Kung University Hospital, the Animal Center of National Cheng Kung University, and the National Center for Genome Medicine (technical/bioinformatics). The authors would like to thank Drs Yi-Chuang Liao and Po-Min Chiang for pathological consultations and Dr Cindy Lee for manuscript editing. This work is supported by the Bi-institutional Collaborative Pancreatic Cancer Research grant from the College of Medicine of National Cheng Kung University to PJL and CSC; and grants MOST102-2320-B-006-050 and MOST104-2320-B-006-028- from the Ministry of Science and Technology, Taiwan, to PHH; MOHW104-TDU-B-211-124-003 from the Ministry of Health and Welfare, Taiwan, to YSS; and MOST 105-2321-B-001-064 from the Team of Excellent Research Program of the Ministry of Science and Technology, Taiwan, to CSC.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    P-H Huang, L-Y Ding, W-Y Hsu, C-S Chen & B-H Chen

  2. Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    P-H Huang, P-J Lu, L-Y Ding, P-C Chu, C-S Chen, Y-S Shan & C-S Chen

  3. Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    P-J Lu, C-C Tsao & Y-S Shan

  4. Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan

    P-C Chu & C-S Chen

  5. Department of Pathology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    C-T Lee

  6. Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Y-S Shan

  7. Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University, Columbus, OH, USA

    C-S Chen

Authors
  1. P-H Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P-J Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L-Y Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P-C Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. W-Y Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C-S Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. C-C Tsao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. B-H Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. C-T Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Y-S Shan
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. C-S Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Y-S Shan or C-S Chen.

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

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, PH., Lu, PJ., Ding, LY. et al. TGFβ promotes mesenchymal phenotype of pancreatic cancer cells, in part, through epigenetic activation of VAV1. Oncogene 36, 2202–2214 (2017). https://doi.org/10.1038/onc.2016.378

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2016.378

This article is cited by

Search

Advanced search

Quick links