Abstract
Overactivation of Wnt-β-catenin signaling, including β-catenin-TCF target gene expression, is a hallmark of colorectal cancer (CRC) development. We identified the immunoglobulin family of cell-adhesion receptors member L1 as a β-catenin-TCF target gene preferentially expressed at the invasive edge of human CRC tissue. L1 can confer enhanced motility and liver metastasis when expressed in CRC cells. This ability of L1-mediated metastasis is exerted by a mechanism involving ezrin and the activation of NF-κB target genes. In this study, we identified the secreted modular calcium-binding matricellular protein-2 (SMOC-2) as a gene activated by L1-ezrin-NF-κB signaling. SMOC-2 is also known as an intestinal stem cell signature gene in mice expressing Lgr5 in cells at the bottom of intestinal crypts. The induction of SMOC-2 expression in L1-expressing CRC cells was necessary for the increase in cell motility, proliferation under stress and liver metastasis conferred by L1. SMOC-2 expression induced a more mesenchymal like phenotype in CRC cells, a decrease in E-cadherin and an increase in Snail by signaling that involves integrin-linked kinase (ILK). SMOC-2 was localized at the bottom of normal human colonic crypts and at increased levels in CRC tissue with preferential expression in invasive areas of the tumor. We found an increase in Lgr5 levels in CRC cells overexpressing L1, p65 or SMOC-2, suggesting that L1-mediated CRC progression involves the acquisition of a stem cell-like phenotype, and that SMOC-2 elevation is necessary for L1-mediated induction of more aggressive/invasive CRC properties.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Clevers H . Wnt/beta-catenin signaling in development and disease. Cell 2006; 127: 469–480.
Polakis P . The many ways of Wnt in cancer. Curr Opin Genet Dev 2007; 17: 45–51.
Conacci-Sorrell M, Zhurinsky J, Ben-Ze’ev A . The cadherin-catenin adhesion system in adhesion, signaling and cancer. J Clin Invest 2002; 109: 987–991.
Brümmendorf T, Kenwrick S, Rathjen FG . Neural cell recognition molecule L1: from cell biology to human hereditary brain malformations. Curr Opin Neurobiol 1998; 8: 87–97.
Hortsch M . Structural and functional evolution of the L1 family: are four adhesion molecules better than one? Mol Cell Neurosci 2000; 15: 1–10.
Conacci-Sorrell ME, Ben-Yedidia T, Shtutman M, Feinstein E, Einat P, Ben-Ze'ev A . Nr-CAM is a target gene of the beta-catenin/LEF-1 pathway in melanoma and colon cancer and its expression enhances motility and confers tumorigenesis. Genes Dev 2002; 16: 2058–2072.
Gavert N, Conacci-Sorrell M, Gast D, Schneider A, Altevogt P, Brabletz T et al. L1, a novel target of beta-catenin signaling, transforms cells and is expressed at the invasive front of colon cancers. J Cell Biol 2005; 168: 633–642.
Gavert N, Sheffer M, Raveh S, Spaderna S, Shtutman M, Brabletz T et al. Expression of L1-CAM and ADAM10 in human colon cancer cells induces metastasis. Cancer Res 2007; 67: 7703–7712.
Gavert N, Shvab A, Sheffer M, Ben-Shmuel A, Haase G, Bakos E et al. c-Kit is suppressed in human colon cancer tissue and contributes to L1-mediated metastasis. Cancer Res 2013; 73: 5754–5763.
Gavert N, Ben-Shmuel A, Lemmon V, Brabletz T, Ben-Ze'ev A . Nuclear factor-kappaB signaling and ezrin are essential for L1-mediated metastasis of colon cancer cells. J Cell Sci 2010; 123: 2135–2143.
Ben-Shmuel A, Shvab A, Gavert N, Brabletz T, Ben-Ze'ev A . Global analysis of L1-transcriptomes identified IGFBP-2 as a target of ezrin and NF-κB signaling that promotes colon cancer progression. Oncogene 2013; 32: 3220–3230.
Munoz J, Stange D, Schepers A, van de Wetering M, Koo B, Itzkovitz S et al. The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent ‘+4’ cell markers. EMBO J 2012; 31: 3079–3091.
Vannahme C, Gosling S, Paulsson M, Maurer P, Hartmann U . Characterization of SMOC-2, a modular extracellular calcium-binding protein. Biochem J 2003; 373: 805–814.
Novinec M, Kovacic L, Skrlj N, Turk V, Lenarcic B . Recombinant human SMOCs produced by in vitro refolding: calcium binding properties and interactions with serum proteins. Protein Expr Purif 2008; 62: 75–82.
Liu P, Lu J, Cardoso W, Vaziri C . The SPARC-related factor SMOC-2 promotes growth factor-induced cyclin D1 expression and DNA synthesis via integrin-linked kinase. Mol Biol Cell 2008; 19: 248–261.
Thiery JP, Acloque H, Huang R, Nieto MA . Epithelial-mesenchymal transitions in development and disease. Cell 2009; 139: 871–890.
Serrano I, McDonald P, Lock F, Muller WJ, Dedhar S . Inactivation of the Hippo tumour suppressor pathway by integrin-linked kinase. Nat Commun 2013; 4: 2976.
Bornstein P, Sage H . Matricellular proteins: extracellular modulators of cell function. Curr Opin Cell Biol 2002; 14: 608–616.
Liu P, Pazin D, Merson R, Albrecht K, Vaziri C . The developmentally-regulated Smoc2 gene is repressed by aryl-hydrocarbon receptor (Ahr) signaling. Gene 2009; 433: 72–80.
Rocnik E, Liu P, Sato K, Walsh K, Vaziri C . The novel SPARC family member SMOC-2 potentiates angiogenic growth factor activity. J Biol Chem 2006; 281: 22855–22864.
Maier S, Paulsson M, Hartmann U . The widely expressed extracellular matrix protein SMOC-2 promotes keratinocyte attachment and migration. Exp Cell Res 2008; 314: 2477–2487.
Milano S, Kwon W, Pereira R, Antonyak M, Cerione R . Characterization of a novel activated Ran GTPase mutant and its ability to induce cellular transformation. J Biol Chem 2012; 287: 24955–24966.
Merlos-Suarez A, Barriga F, Jung P, Iglesias M, Cespedes MV, Rossell D et al. The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse. Cell Stem Cell 2011; 8: 511–524.
Uchida H, Yamazaki K, Fukuma M, Yamada T, Hayashida T et al. Overexpression of leucine-rich repeat-containing G protein-coupled receptor 5 in colorectal cancer. Cancer Sci 2010; 101: 1731–1737.
Schwitalla S, Fingerle A, Cammareri P, Nebelsiek T, Goktuna S et al. Intestinal tumorigenesis initiated by dedifferentiation and acquisition of stem-cell-like properties. Cell 2013; 152: 25–38.
Pattabiraman D, Weinberg R . Tackling the cancer stem cells—what challenges do they pose? Nature Rev Drug Discov 2014; 13: 497–512.
Brabletz T, Jung A, Spaderna S, Hlubek F, Kirchner T . Opinion: migrating cancer stem cells—an integrated concept of malignant tumor progression. Nat Rev Cancer 2005; 5: 744–749.
Brabletz T . EMT and MET in metastasis: where are the cancer stem cells? Can Cell 2012; 22: 699–701.
Gavert N, Vivanti A, Hazin J, Brabletz T, Ben-Ze’ev A . L1-mediated colon cancer metastasis does not require changes in EMT and cancer stem cell markers. Mol Can Res 2011; 9: 14–24.
Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T et al. CD133 expression is not restricted to stem cells, and both CD133+ and CD133-metastatic colon cancer cells initiate tumors. J Clin Invest 2008; 118: 2111–2120.
Yan Z, Yin H, Wang R, Wu D, Sun W et al. Overexpression of integrin-linked kinase (ILK) promotes migration and invasion of colorectal cancer cells by inducing epithelial-mesenchymal transition via NF-κB signaling. Acta Histochem 2014; 116: 527–533.
Simons B, Clevers H . Stem cell self-renewal in intestinal crypt. Exp Cell Res 2011; 317: 2719–2724.
Acknowledgements
We thank Drs C Vaziri, U Hartmann, B Lenarcic, S Milano, R Cerione and P McDonald for reagents. This study was supported by grants from the Israel Cancer Research Fund (ICRF) and from the Israel Science Foundation (ISF).
Author information
Authors and Affiliations
Corresponding author
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
About this article
Cite this article
Shvab, A., Haase, G., Ben-Shmuel, A. et al. Induction of the intestinal stem cell signature gene SMOC-2 is required for L1-mediated colon cancer progression. Oncogene 35, 549–557 (2016). https://doi.org/10.1038/onc.2015.127
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2015.127
This article is cited by
-
SMOC2 promotes aggressive behavior of fibroblast-like synoviocytes in rheumatoid arthritis through transcriptional and post-transcriptional regulating MYO1C
Cell Death & Disease (2022)
-
SMOC2 promotes an epithelial-mesenchymal transition and a pro-metastatic phenotype in epithelial cells of renal cell carcinoma origin
Cell Death & Disease (2022)
-
A positive feedback loop between Periostin and TGFβ1 induces and maintains the stemness of hepatocellular carcinoma cells via AP-2α activation
Journal of Experimental & Clinical Cancer Research (2021)
-
The epigenetic regulator Mll1 is required for Wnt-driven intestinal tumorigenesis and cancer stemness
Nature Communications (2020)
-
SMOC2, an intestinal stem cell marker, is an independent prognostic marker associated with better survival in colorectal cancers
Scientific Reports (2020)