Digestive Diseases and Sciences

, Volume 64, Issue 10, pp 2830–2842 | Cite as

Comprehensive Analysis of the Canonical and Non-canonical Wnt Signaling Pathways in Gastric Cancer

  • Le Wang
  • Hao Wang
  • Xianglong Duan
  • Penggao DaiEmail author
  • Jianping LiEmail author
Original Article



Previous studies showed that dysregulation of Wnt signaling by gene mutation and abnormal gene expression is one of the causative factors for gastric cancer (GC). So far, a systematic and comprehensive analysis of gene mutation, gene expression, and DNA methylation profiles of the Wnt pathway associated with gastric carcinogenesis, however, has not yet been reported.


To this end, we investigated all the above-mentioned genetic alterations associated with the canonical and non-canonical Wnt pathways in GC tumors, in order to understand the molecular mechanism underlying gastric carcinogenesis.


The information on gene mutations and expression was obtained from data resources, such as TCGA, GSEA, and TCGA-STAD, and was analyzed with the cBioPortal platform. We also performed in vitro analysis on DDK2 gene, a Wnt inhibitor, to characterize its role in GC tumor cells.


We found that gene mutations of 43 Wnt genes and abnormal expression of 13 Wnt genes occurred at a high frequency in GC tumors, and gene amplification and deletion are the major mutation types. Clusters of DNA methylation associated with Wnt signaling genes and GC tumors were also revealed, and a significant increase in β-catenin activity was found in the hypermethylated group of GC tumors. In addition, overexpression of DKK2 gene significantly inhibited multiple biological processes of the GC cells, including their growth, clonal forming, migration, and invasion ability, and induced apoptosis of the GC cells.


Our current study suggested that gene mutation, abnormal gene expression, and altered DNA methylation profiles associated with the Wnt signaling may play an important role in gastric carcinogenesis, and DKK2 gene may act as a tumor suppressor in gastric cells.


Wnt signaling pathway Gene mutation mRNA expression DNA methylation Gastric cancer 



This study was funded by the National Natural Science Foundation of China (Grant No. 81760441).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10620_2019_5606_MOESM1_ESM.doc (280 kb)
Supplementary material 1 (DOC 279 kb)
10620_2019_5606_MOESM2_ESM.xlsx (38 kb)
Supplementary material 2 (XLSX 37 kb)


  1. 1.
    Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015. CA Cancer J Clin. 2016;66:115–132.CrossRefGoogle Scholar
  2. 2.
    Suh YS, Yang HK. Screening and early detection of gastric cancer: east versus west. Surg Clin N Am. 2015;95:1053–1066.CrossRefGoogle Scholar
  3. 3.
    Kanda M, Oya H, Nomoto S, et al. Diversity of clinical implication of B-cell translocation gene 1 expression by histopathologic and anatomic subtypes of gastric cancer. Dig Dis Sci. 2015;60:1256–1264. Scholar
  4. 4.
    Huang GL, Luo Q, Rui G, et al. Oncogenic activity of retinoic acid receptor gamma is exhibited through activation of the Akt/NF-kappaB and Wnt/beta-catenin pathways in cholangiocarcinoma. Mol Cell Biol. 2013;33:3416–3425.CrossRefGoogle Scholar
  5. 5.
    Hiyama T, Haruma K, Kitadai Y, et al. K-ras mutation in helicobacter pylori-associated chronic gastritis in patients with and without gastric cancer. Int J Cancer. 2002;97:562–566.CrossRefGoogle Scholar
  6. 6.
    Chiurillo MA. Role of the Wnt/beta-catenin pathway in gastric cancer: an in-depth literature review. World J Exp Med. 2015;5:84–102.CrossRefGoogle Scholar
  7. 7.
    Wang K, Yuen ST, Xu J, et al. Whole-genome sequencing and comprehensive molecular profiling identify new driver mutations in gastric cancer. Nat Genet. 2014;46:573–582.CrossRefGoogle Scholar
  8. 8.
    Radulescu S, Ridgway RA, Cordero J, et al. Acute WNT signalling activation perturbs differentiation within the adult stomach and rapidly leads to tumour formation. Oncogene. 2013;32:2048–2057.CrossRefGoogle Scholar
  9. 9.
    Yamaguchi T, Yanagisawa K, Sugiyama R, et al. NKX2-1/TITF1/TTF-1-Induced ROR1 is required to sustain EGFR survival signaling in lung adenocarcinoma. Cancer Cell. 2012;21:348–361.CrossRefGoogle Scholar
  10. 10.
    Zhang S, Chen L, Cui B, et al. ROR1 is expressed in human breast cancer and associated with enhanced tumor-cell growth. PLoS ONE. 2012;7:e31127.CrossRefGoogle Scholar
  11. 11.
    Zhan T, Rindtorff N, Boutros M. Wnt signaling in cancer. Oncogene. 2017;36:1461–1473.CrossRefGoogle Scholar
  12. 12.
    Papkoff J, Brown AM, Varmus HE. The int-1 proto-oncogene products are glycoproteins that appear to enter the secretory pathway. Mol Cell Biol. 1987;7:3978–3984.CrossRefGoogle Scholar
  13. 13.
    He TC, Sparks AB, Rago C, et al. Identification of c-MYC as a target of the APC pathway. Science. 1998;281:1509–1512.CrossRefGoogle Scholar
  14. 14.
    Shtutman M, Zhurinsky J, Simcha I, et al. The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc Natl Acad Sci USA. 1999;96:5522–5527.CrossRefGoogle Scholar
  15. 15.
    Koh TJ, Bulitta CJ, Fleming JV, Dockray GJ, Varro A, Wang TC. Gastrin is a target of the beta-catenin/TCF-4 growth-signaling pathway in a model of intestinal polyposis. J Clin Investig. 2000;106:533–539.CrossRefGoogle Scholar
  16. 16.
    Kolligs FT, Nieman MT, Winer I, et al. ITF-2, a downstream target of the Wnt/TCF pathway, is activated in human cancers with beta-catenin defects and promotes neoplastic transformation. Cancer Cell. 2002;1:145–155.CrossRefGoogle Scholar
  17. 17.
    Sasai N, Nakazawa Y, Haraguchi T, Sasai Y. The neurotrophin-receptor-related protein NRH1 is essential for convergent extension movements. Nat Cell Biol. 2004;6:741–748.CrossRefGoogle Scholar
  18. 18.
    Lu W, Yamamoto V, Ortega B, Baltimore D. Mammalian Ryk is a Wnt coreceptor required for stimulation of neurite outgrowth. Cell. 2004;119:97–108.CrossRefGoogle Scholar
  19. 19.
    Lu X, Borchers AG, Jolicoeur C, Rayburn H, Baker JC, Tessier-Lavigne M. PTK7/CCK-4 is a novel regulator of planar cell polarity in vertebrates. Nature. 2004;430:93–98.CrossRefGoogle Scholar
  20. 20.
    Nishita M, Yoo SK, Nomachi A, et al. Filopodia formation mediated by receptor tyrosine kinase Ror2 is required for Wnt5a-induced cell migration. J Cell Biol. 2006;175:555–562.CrossRefGoogle Scholar
  21. 21.
    Marlow F, Topczewski J, Sepich D, Solnica-Krezel L. Zebrafish Rho kinase 2 acts downstream of Wnt11 to mediate cell polarity and effective convergence and extension movements. Curr Biol. 2002;12:876–884.CrossRefGoogle Scholar
  22. 22.
    Li L, Yuan H, Xie W, et al. Dishevelled proteins lead to two signaling pathways. Regulation of LEF-1 and c-Jun N-terminal kinase in mammalian cells. J Biol Chem. 1999;274:129–134.CrossRefGoogle Scholar
  23. 23.
    Sheldahl LC, Slusarski DC, Pandur P, Miller JR, Kuhl M, Moon RT. Dishevelled activates Ca2+ flux, PKC, and CamKII in vertebrate embryos. J Cell Biol. 2003;161:769–777.CrossRefGoogle Scholar
  24. 24.
    Kuhl M, Sheldahl LC, Malbon CC, Moon RT. Ca(2+)/calmodulin-dependent protein kinase II is stimulated by Wnt and Frizzled homologs and promotes ventral cell fates in Xenopus. J Biol Chem. 2000;275:12701–12711.CrossRefGoogle Scholar
  25. 25.
    Zhu J, Zhang S, Gu L, Di W. Epigenetic silencing of DKK2 and Wnt signal pathway components in human ovarian carcinoma. Carcinogenesis. 2012;33:2334–2343.CrossRefGoogle Scholar
  26. 26.
    Chalmers ZR, Connelly CF, Fabrizio D, et al. Analysis of 100,000 human cancer genomes reveals the landscape of tumor mutational burden. Genome Med. 2017;9:34.CrossRefGoogle Scholar
  27. 27.
    Moustakas A, Souchelnytskyi S, Heldin CH. Smad regulation in TGF-beta signal transduction. J Cell Sci. 2001;114:4359–4369.Google Scholar
  28. 28.
    Silva AL, Dawson SN, Arends MJ, et al. Boosting Wnt activity during colorectal cancer progression through selective hypermethylation of Wnt signaling antagonists. BMC Cancer. 2014;14:891.CrossRefGoogle Scholar
  29. 29.
    Cancer Genome Atlas Research Network. Comprehensive molecular characterization of gastric adenocarcinoma. Nature. 2014;513:202–209. Scholar
  30. 30.
    Toyota M, Ahuja N, Ohe-Toyota M, Herman JG, Baylin SB, Issa JP. CpG island methylator phenotype in colorectal cancer. Proc Natl Acad Sci USA. 1999;96:8681–8686.CrossRefGoogle Scholar
  31. 31.
    Duchartre Y, Kim YM, Kahn M. The Wnt signaling pathway in cancer. Crit Rev Oncol Hematol. 2016;99:141–149.CrossRefGoogle Scholar
  32. 32.
    Wang H, Duan XL, Qi XL, et al. Concurrent hypermethylation of SFRP2 and DKK2 activates the Wnt/beta-catenin pathway and is associated with poor prognosis in patients with gastric cancer. Mol Cells. 2017;40:45–53.CrossRefGoogle Scholar
  33. 33.
    Ooi CH, Ivanova T, Wu J, et al. Oncogenic pathway combinations predict clinical prognosis in gastric cancer. PLoS Genet. 2009;5:e1000676.CrossRefGoogle Scholar
  34. 34.
    Yoda Y, Takeshima H, Niwa T, et al. Integrated analysis of cancer-related pathways affected by genetic and epigenetic alterations in gastric cancer. Gastric Cancer. 2015;18:65–76.CrossRefGoogle Scholar
  35. 35.
    Ji J, Feng X, Shi M, et al. Rac1 is correlated with aggressiveness and a potential therapeutic target for gastric cancer. Int J Oncol. 2015;46:1343–1353.CrossRefGoogle Scholar
  36. 36.
    Uematsu K, He B, You L, Xu Z, McCormick F, Jablons DM. Activation of the Wnt pathway in non small cell lung cancer: evidence of dishevelled overexpression. Oncogene. 2003;22:7218–7221.CrossRefGoogle Scholar
  37. 37.
    Shan YS, Hsu HP, Lai MD, et al. Cyclin D1 overexpression correlates with poor tumor differentiation and prognosis in gastric cancer. Oncol Lett. 2017;14:4517–4526.CrossRefGoogle Scholar
  38. 38.
    Sato H, Suzuki H, Toyota M, et al. Frequent epigenetic inactivation of DICKKOPF family genes in human gastrointestinal tumors. Carcinogenesis. 2007;28:2459–2466.CrossRefGoogle Scholar
  39. 39.
    Nojima M, Suzuki H, Toyota M, et al. Frequent epigenetic inactivation of SFRP genes and constitutive activation of Wnt signaling in gastric cancer. Oncogene. 2007;26:4699–4713.CrossRefGoogle Scholar
  40. 40.
    Guo Y, Guo W, Chen Z, Kuang G, Yang Z, Dong Z. Hypermethylation and aberrant expression of Wnt-antagonist family genes in gastric cardia adenocarcinoma. Neoplasma. 2011;58:110–117.CrossRefGoogle Scholar
  41. 41.
    Zhao Z, Liu W, Liu J, Wang J, Luo B. The effect of EBV on WIF1, NLK, and APC gene methylation and expression in gastric carcinoma and nasopharyngeal cancer. J Med Virol. 2017;89:1844–1851.CrossRefGoogle Scholar
  42. 42.
    Deng J, Liang H, Zhang R, et al. Methylated CpG site count of dapper homolog 1 (DACT1) promoter prediction the poor survival of gastric cancer. Am J Cancer Res. 2014;4:518–527.Google Scholar
  43. 43.
    Yu Y, Yan W, Liu X, et al. DACT2 is frequently methylated in human gastric cancer and methylation of DACT2 activated Wnt signaling. Am J Cancer Res. 2014;4:710–724.Google Scholar
  44. 44.
    Lee HK, Chaboub LS, Zhu W, et al. Daam2-PIP5K is a regulatory pathway for Wnt signaling and therapeutic target for remyelination in the CNS. Neuron. 2015;85:1227–1243.CrossRefGoogle Scholar
  45. 45.
    Rodriguez N, Yang J, Hasselblatt K, et al. Casein kinase I epsilon interacts with mitochondrial proteins for the growth and survival of human ovarian cancer cells. EMBO Mol Med. 2012;4:952–963.CrossRefGoogle Scholar
  46. 46.
    Polakis P. The many ways of Wnt in cancer. Curr Opin Genet Dev. 2007;17:45–51.CrossRefGoogle Scholar
  47. 47.
    Peters JM, McKay RM, McKay JP, Graff JM. Casein kinase I transduces Wnt signals. Nature. 1999;401:345–350.CrossRefGoogle Scholar
  48. 48.
    Surana R, Sikka S, Cai W, et al. Secreted frizzled related proteins: implications in cancers. Biochim Biophys Acta. 2014;1845:53–65.Google Scholar
  49. 49.
    Niehrs C. Function and biological roles of the Dickkopf family of Wnt modulators. Oncogene. 2006;25:7469–7481.CrossRefGoogle Scholar
  50. 50.
    Maehata T, Taniguchi H, Yamamoto H, et al. Transcriptional silencing of Dickkopf gene family by CpG island hypermethylation in human gastrointestinal cancer. World J Gastroenterol. 2008;14:2702–2714.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Oncology, The First Affiliated Hospital of Medical CollegeXi’an Jiaotong UniversityXi’anChina
  2. 2.National Engineering Research Center for Miniaturized Detection Systems, School of Life SciencesNorthwest UniversityXi’anChina
  3. 3.The Second Department of General SurgeryShaanxi Provincial People’s HospitalXi’anChina
  4. 4.Department of General SurgeryThe First Hospital of YulinShaanxiChina
  5. 5.Department of Otolaryngology Xi’an No.4 HospitalXi’anChina

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