Concurrent Hypermethylation of SFRP2 and DKK2 Activates the Wnt/β-Catenin Pathway and Is Associated with Poor Prognosis in Patients with Gastric Cancer

  • Wang, Hao (National Engineering Research Center for Miniaturized Detection Systems, School of Life Sciences, Northwest University) ;
  • Duan, Xiang-Long (Second Department of General Surgery, Shaanxi Provincial People's Hospital) ;
  • Qi, Xiao-Li (National Engineering Research Center for Miniaturized Detection Systems, School of Life Sciences, Northwest University) ;
  • Meng, Lei (National Engineering Research Center for Miniaturized Detection Systems, School of Life Sciences, Northwest University) ;
  • Xu, Yi-Song (National Engineering Research Center for Miniaturized Detection Systems, School of Life Sciences, Northwest University) ;
  • Wu, Tong (National Engineering Research Center for Miniaturized Detection Systems, School of Life Sciences, Northwest University) ;
  • Dai, Peng-Gao (National Engineering Research Center for Miniaturized Detection Systems, School of Life Sciences, Northwest University)
  • Received : 2016.10.18
  • Accepted : 2016.12.20
  • Published : 2017.01.31


Aberrant hypermethylation of Wnt antagonists has been observed in gastric cancer. A number of studies have focused on the hypermethylation of a single Wnt antagonist and its role in regulating the activation of signaling. However, how the Wnt antagonists interacted to regulate the signaling pathway has not been reported. In the present study, we systematically investigated the methylation of some Wnt antagonist genes (SFRP2, SFRP4, SFRP5, DKK1, DKK2, and APC) and their regulatory role in carcinogenesis. We found that aberrant promoter methylation of SFRP2, SFRP4, DKK1, and DKK2 was significantly increased in gastric cancer. Moreover, concurrent hypermethylation of SFRP2 and DKK2 was observed in gastric cancer and this was significantly associated with increased expression of ${\beta}-catenin$, indicating that the joint inactivation of these two genes promoted the activation of the Wnt signaling pathway. Further analysis using a multivariate Cox proportional hazards model showed that DKK2 methylation was an independent prognostic factor for poor overall survival, and the predictive value was markedly enhanced when the combined methylation status of SFRP2 and DKK2 was considered. In addition, the methylation level of SFRP4 and DKK2 was correlated with the patient's age and tumor differentiation, respectively. In conclusion, epigenetic silencing of Wnt antagonists was associated with gastric carcinogenesis, and concurrent hypermethylation of SFRP2 and DKK2 could be a potential marker for a prognosis of poor overall survival.


${\beta}-catenin$;DNA methylation;gastric cancer;prognosis;Wnt antagonists


Supported by : National Natural Science Foundation of China


  1. Behrens, J., von Kries, J.P., Kuhl, M., Bruhn, L., Wedlich, D., Grosschedl, R., and Birchmeier, W. (1996). Functional interaction of beta-catenin with the transcription factor LEF-1. Nature 382, 638-642.
  2. Cadigan, K.M., and Nusse, R. (1997). Wnt signaling: a common theme in animal development. Genes Dev. 11, 3286-3305.
  3. Cheng, Y.Y., Yu, J., Wong, Y.P., Man, E.P., To, K.F., Jin, V.X., Li, J., Tao, Q., Sung, J.J., Chan, F.K., et al. (2007). Frequent epigenetic inactivation of secreted frizzled-related protein 2 (SFRP2) by promoter methylation in human gastric cancer. Br. J. Cancer 97, 895-901.
  4. Clements, W.M., Wang, J., Sarnaik, A., Kim, O.J., MacDonald, J., Fenoglio-Preiser, C., Groden, J., and Lowy, A.M. (2002). beta-Catenin mutation is a frequent cause of Wnt pathway activation in gastric cancer. Cancer Res. 62, 3503-3506.
  5. Ferlay, J., Shin, H.R., Bray, F., Forman, D., Mathers, C., and Parkin, D.M. (2010). Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int. J. Cancer 127, 2893-2917.
  6. Ford, C.E., Jary, E., Ma, S.S., Nixdorf, S., Heinzelmann-Schwarz, V.A., and Ward, R.L. (2013). The Wnt gatekeeper SFRP4 modulates EMT, cell migration and downstream Wnt signalling in serous ovarian cancer cells. PloS one 8, e54362.
  7. Giles, R.H., van Es, J.H., and Clevers, H. (2003). Caught up in a Wnt storm: Wnt signaling in cancer. Biochim. Biophys. Acta 1653, 1-24.
  8. Gonzalez-Sancho, J.M., Brennan, K.R., Castelo-Soccio, L.A., and Brown, A.M. (2004). Wnt proteins induce dishevelled phosphorylation via an LRP5/6- independent mechanism, irrespective of their ability to stabilize beta-catenin. Mol. Cell. Biol. 24, 4757-4768.
  9. Hirata, H., Hinoda, Y., Nakajima, K., Kawamoto, K., Kikuno, N., Kawakami, K., Yamamura, S., Ueno, K., Majid, S., Saini, S., et al. (2009). Wnt antagonist gene DKK2 is epigenetically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways. Clin. Cancer Res. 15, 5678-5687.
  10. Hosoya, K., Yamashita, S., Ando, T., Nakajima, T., Itoh, F., and Ushijima, T. (2009). Adenomatous polyposis coli 1A is likely to be methylated as a passenger in human gastric carcinogenesis. Cancer Lett. 285, 182-189.
  11. Kikuchi, A., Yamamoto, H., Sato, A., and Matsumoto, S. (2012). Wnt5a: its signalling, functions and implication in diseases. Acta Physiol. 204, 17-33.
  12. Li, S., Wang, C., Liu, X., Hua, S., and Liu, X. (2015). The roles of AXIN2 in tumorigenesis and epigenetic regulation. Familial Cancer 14, 325-331.
  13. Liu, G., Bafico, A., and Aaronson, S.A. (2005). The mechanism of endogenous receptor activation functionally distinguishes prototype canonical and noncanonical Wnts. Mol. Cell. Biol. 25, 3475-3482.
  14. Lu, W., Yamamoto, V., Ortega, B., and Baltimore, D. (2004a). Mammalian Ryk is a Wnt coreceptor required for stimulation of neurite outgrowth. Cell 119, 97-108.
  15. Lu, X., Borchers, A.G., Jolicoeur, C., Rayburn, H., Baker, J.C., and Tessier-Lavigne, M. (2004b). PTK7/CCK-4 is a novel regulator of planar cell polarity in vertebrates. Nature 430, 93-98.
  16. Mann, B., Gelos, M., Siedow, A., Hanski, M.L., Gratchev, A., Ilyas, M., Bodmer, W.F., Moyer, M.P., Riecken, E.O., Buhr, H.J., et al. (1999). Target genes of beta-catenin-T cell-factor/lymphoid-enhancer-factor signaling in human colorectal carcinomas. Proc. Natl. Acad. Sci. USA 96, 1603-1608.
  17. Niehrs, C. (2006). Function and biological roles of the Dickkopf family of Wnt modulators. Oncogene 25, 7469-7481.
  18. Nojima, M., Suzuki, H., Toyota, M., Watanabe, Y., Maruyama, R., Sasaki, S., Sasaki, Y., Mita, H., Nishikawa, N., Yamaguchi, K., et al. (2007). Frequent epigenetic inactivation of SFRP genes and constitutive activation of Wnt signaling in gastric cancer. Oncogene 26, 4699-4713.
  19. Oishi, I., Suzuki, H., Onishi, N., Takada, R., Kani, S., Ohkawara, B., Koshida, I., Suzuki, K., Yamada, G., Schwabe, G.C., et al. (2003). The receptor tyrosine kinase Ror2 is involved in non-canonical Wnt5a/JNK signalling pathway. Genes Cells 8, 645-654.
  20. Ooi, C.H., Ivanova, T., Wu, J., Lee, M., Tan, I.B., Tao, J., Ward, L., Koo, J.H., Gopalakrishnan, V., Zhu, Y., et al. (2009). Oncogenic pathway combinations predict clinical prognosis in gastric cancer. PLoS Genet. 5, e1000676.
  21. Sato, H., Suzuki, H., Toyota, M., Nojima, M., Maruyama, R., Sasaki, S., Takagi, H., Sogabe, Y., Sasaki, Y., Idogawa, M., et al. (2007). Frequent epigenetic inactivation of DICKKOPF family genes in human gastrointestinal tumors. Carcinogenesis 28, 2459-2466.
  22. Silva, A.L., Dawson, S.N., Arends, M.J., Guttula, K., Hall, N., Cameron, E.A., Huang, T.H., Brenton, J.D., Tavare, S., Bienz, M., et al. (2014). Boosting Wnt activity during colorectal cancer progression through selective hypermethylation of Wnt signaling antagonists. BMC Cancer 14, 891.
  23. Tsuchiya, T., Tamura, G., Sato, K., Endoh, Y., Sakata, K., Jin, Z., Motoyama, T., Usuba, O., Kimura, W., Nishizuka, S., et al. (2000). Distinct methylation patterns of two APC gene promoters in normal and cancerous gastric epithelia. Oncogene 19, 3642-3646.
  24. Veeck, J., Noetzel, E., Bektas, N., Jost, E., Hartmann, A., Knuchel, R., and Dahl, E. (2008). Promoter hypermethylation of the SFRP2 gene is a high-frequent alteration and tumor-specific epigenetic marker in human breast cancer. Mol. Cancer 7, 83.
  25. Wang, Z.K., Liu, J., Liu, C., Wang, F.Y., Chen, C.Y., and Zhang, X.H. (2012). Hypermethylation of adenomatous polyposis coli gene promoter is associated with novel Wnt signaling pathway in gastric adenomas. J. Gastroenterol. Hepatol. 27, 1629-1634.
  26. Woo, D.K., Kim, H.S., Lee, H.S., Kang, Y.H., Yang, H.K., and Kim, W.H. (2001). Altered expression and mutation of beta-catenin gene in gastric carcinomas and cell lines. Int. J. Cancer 95, 108-113.<108::AID-IJC1019>3.0.CO;2-#
  27. Yoda, Y., Takeshima, H., Niwa, T., Kim, J.G., Ando, T., Kushima, R., Sugiyama, T., Katai, H., Noshiro, H., and Ushijima, T. (2015). Integrated analysis of cancer-related pathways affected by genetic and epigenetic alterations in gastric cancer. Gastric Cancer 18, 65-76.
  28. Yu, J., Tao, Q., Cheng, Y.Y., Lee, K.Y., Ng, S.S., Cheung, K.F., Tian, L., Rha, S.Y., Neumann, U., Rocken, C., et al. (2009). Promoter methylation of the Wnt/beta-catenin signaling antagonist Dkk-3 is associated with poor survival in gastric cancer. Cancer 115, 49-60.

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