Abstract
A large proportion of malignant cancers of the stomach are gastric adenocarcinoma type. In spite of many studies, the molecular basis for this cancer is still unclear. Deregulated cell proliferative signaling via Wnt/β-catenin and Hedgehog pathways is considered important in the pathogenesis of many cancers including the gastric cancer. Recent studies identified ZnRF3 protein, which is a E3-ubiquitin ligase and which is either deleted or mutated in cancers, to inhibit Wnt signaling. However, the significance of ZnRF3 in the control of gastric cancer and whether it also regulates Hedgehog signaling pathway, is not known. In the present study, we assessed the expression of ZnRF3 in gastric tumors and paracancerous tissues from 58 patients (44 male and 14 female) of different ages and related this to patient survival. We observed a clear relationship between ZnRF3 expression in paracancerous tissue and tumor size. Also, ZnRF3 expression was much higher in tumors from aged patients. Male patients showed higher mortality than the females. Mechanistic studies using normal gastric cells (GES1) and gastric cancer cells (MGC-803) infected with either AdZnRF3 or AdGFP viral vectors, revealed that ZnRF3 overexpression causes significantly more apoptosis and lowered proliferation of cancer cells. ZnRF3 overexpression led to greatly reduced levels of Lgr5, a component of Wnt signaling and also Gli1, a component of Hedgehog signaling. Thus, ZnRF3 negatively influences both the Wnt and Hedgehog proliferative pathways, and probably this way it negatively regulates cancer progression. These results suggest the importance of normal ZnRF3 function in checking the progression of cancer cell growth and indicate that a lack of this protein can lead to poorer clinical outcomes for gastric cancer patients.
Similar content being viewed by others
References
Smith, M. G., Hold, G. L., Tahara, E., & El-Omar, E. M. (2006). Cellular and molecular aspects of gastric cancer. World Journal of Gastroenterology, 12, 2979–2990.
Ushijima, T., & Sasako, M. (2004). Focus on gastric cancer. Cancer Cell, 5, 121–125.
Cho, L. Y., Yang, J. J., Ko, K. P., Ma, S. H., Shin, A., Choi, B. Y., et al. (2012). Genetic susceptibility factors on genes involved in the steroid hormone biosynthesis pathway and progesterone receptor for gastric cancer risk. PLoS ONE, 7, e47603.
Clevers, H., & Nusse, R. (2012). Wnt/beta-catenin signaling and disease. Cell, 149, 1192–1205.
Zhou, Y., Lan, J., Wang, W., Shi, Q., Lan, Y., Cheng, Z., & Guan, H. (2013). Znrf3 acts as a tumour suppressor by the Wnt signalling pathway in human gastric adenocarcinoma. Journal of Molecular Histology, 44, 555–563.
Lowy, A. M., Clements, W. M., Bishop, J., Kong, L., Bonney, T., Sisco, K., et al. (2006). Beta-catenin/Wnt signaling regulates expression of the membrane type 3 matrix metalloproteinase in gastric cancer. Cancer Research, 66, 4734–4741.
Clevers, H. (2006). Wnt/beta-catenin signaling in development and disease. Cell, 127, 469–480.
Macdonald, B. T., Tamai, K., & He, X. (2009). Wnt/beta-catenin signaling: Components, mechanisms, and diseases. Developmental Cell, 17, 9–26.
Pecina-Slaus, N. (2010). Wnt signal transduction pathway and apoptosis: A review. Cancer Cell International, 10, 22.
Grossmann, T. N., Yeh, J. T., Bowman, B. R., Chu, Q., Moellering, R. E., & Verdine, G. L. (2012). Inhibition of oncogenic Wnt signaling through direct targeting of beta-catenin. Proceedings of the National Academy of Sciences USA, 109, 17942–17947.
Schepers, A., & Clevers, H. (2012). Wnt signaling, stem cells, and cancer of the gastrointestinal tract. Cold Spring Harbor Perspectives in Biology, 4, a007989.
Kim, K. A., Wagle, M., Tran, K., Zhan, X., Dixon, M. A., Liu, S., et al. (2008). R-spondin family members regulate the Wnt pathway by a common mechanism. Molecular Biology of the Cell, 19, 2588–2596.
Kazanskaya, O., Ohkawara, B., Heroult, M., Wu, W., Maltry, N., Augustin, H. G., & Niehrs, C. (2008). The Wnt signaling regulator R-spondin 3 promotes angioblast and vascular development. Development, 135, 3655–3664.
Parma, P., Radi, O., Vidal, V., Chaboissier, M. C., Dellambra, E., Valentini, S., et al. (2006). R-spondin1 is essential in sex determination, skin differentiation and malignancy. Nature Genetics, 38, 1304–1309.
Ootani, A., Li, X., Sangiorgi, E., Ho, Q. T., Ueno, H., Toda, S., et al. (2009). Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche. Nature Medicine, 15, 701–706.
Carmon, K. S., Gong, X., Lin, Q., Thomas, A., & Liu, Q. (2011). R-spondins function as ligands of the orphan receptors lgr4 and lgr5 to regulate Wnt/beta-catenin signaling. Proceedings of the National Academy of Sciences USA, 108, 11452–11457.
de Lau, W., Barker, N., Low, T. Y., Koo, B. K., Li, V. S., Teunissen, H., et al. (2011). Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling. Nature, 476, 293–297.
Hao, H. X., Xie, Y., Zhang, Y., Charlat, O., Oster, E., Avello, M., et al. (2012). Znrf3 promotes Wnt receptor turnover in an R-spondin-sensitive manner. Nature, 485, 195–200.
de Lau, W., Peng, W. C., Gros, P., & Clevers, H. (2014). The R-spondin/lgr5/rnf43 module: Regulator of Wnt signal strength. Genes & Development, 28, 305–316.
Peng, W. C., de Lau, W., Madoori, P. K., Forneris, F., Granneman, J. C., Clevers, H., & Gros, P. (2013). Structures of Wnt-antagonist znrf3 and its complex with R-spondin 1 and implications for signaling. PLoS ONE, 8, e83110.
Sun, Y. (2006). E3 ubiquitin ligases as cancer targets and biomarkers. Neoplasia, 8, 645–654.
Wolpin, B. M., Rizzato, C., Kraft, P., Kooperberg, C., Petersen, G. M., Wang, Z., et al. (2014). Genome-wide association study identifies multiple susceptibility loci for pancreatic cancer. Nature Genetics, 46, 994–1000.
Infante, P., Canettieri, G., Gulino, A., & di Marcotullio, L. (2014). Yin-yang strands of pcaf/hedgehog axis in cancer control. Trends in Molecular Medicine, 20, 416–418.
Remmele, W., & Stegner, H. E. (1987). Recommendation for uniform definition of an immunoreactive score (irs) for immunohistochemical estrogen receptor detection (er-ica) in breast cancer tissue. Der Pathologe, 8, 138–140.
Brenner, H., Rothenbacher, D., & Arndt, V. (2009). Epidemiology of stomach cancer. Methods in Molecular Biology, 472, 467–477.
Amakye, D., Jagani, Z., & Dorsch, M. (2013). Unraveling the therapeutic potential of the hedgehog pathway in cancer. Nature Medicine, 19, 1410–1422.
Mazza, D., Infante, P., Colicchia, V., Greco, A., Alfonsi, R., Siler, M., et al. (2013). Pcaf ubiquitin ligase activity inhibits hedgehog/gli1 signaling in p53-dependent response to genotoxic stress. Cell Death and Differentiation, 20, 1688–1697.
Acknowledgments
Grants (Nos. 81272698, 81101883, 81172368) from the National Nature Science Foundation of China. A Grant (No. 20130206) from the Special Scientific Research Foundation of Health Sector from the National Health and Family Planning Commission of China. A Grant form the capital health research and development of special (No. 2011-5001-01). A Grant form Major Science and Technology Progect of “National Significant New Drug Creation” from the Major Science and Technology of China (No. 2011ZX09307-001-05).
Author information
Authors and Affiliations
Corresponding author
Additional information
Hongzhen Qin, Aizhen Cai and Hongqing Xi have contributed equally to this study, should be considered as co-first authors.
Rights and permissions
About this article
Cite this article
Qin, H., Cai, A., Xi, H. et al. ZnRF3 Induces Apoptosis of Gastric Cancer Cells by Antagonizing Wnt and Hedgehog Signaling. Cell Biochem Biophys 73, 361–367 (2015). https://doi.org/10.1007/s12013-015-0607-7
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12013-015-0607-7