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Blocking Wnt signaling by SFRP-like molecules inhibits in vivo cell proliferation and tumor growth in cells carrying active β-catenin

Oncogene volume 30, pages 423–433 (2011)Cite this article

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Abstract

Constitutive activation of Wnt/β-catenin signaling in cancer results from mutations in pathway components, which frequently coexist with autocrine Wnt signaling or epigenetic silencing of extracellular Wnt antagonists. Among the extracellular Wnt inhibitors, the secreted frizzled-related proteins (SFRPs) are decoy receptors that contain soluble Wnt-binding frizzled domains. In addition to SFRPs, other endogenous molecules harboring frizzled motifs bind to and inhibit Wnt signaling. One of such molecules is V3Nter, a soluble SFRP-like frizzled polypeptide that binds to Wnt3a and inhibits Wnt signaling and expression of the β-catenin target genes cyclin D1 and c-myc. V3Nter is derived from the cell surface extracellular matrix component collagen XVIII. Here, we used HCT116 human colon cancer cells carrying the ΔS45 activating mutation in one of the alleles of β-catenin to show that V3Nter and SFRP-1 decrease baseline and Wnt3a-induced β-catenin stabilization. Consequently, V3Nter reduces the growth of human colorectal cancer xenografts by specifically controlling cell proliferation and cell cycle progression, without affecting angiogenesis or apoptosis, as shown by decreased [3H]-thymidine (in vitro) or BrdU (in vivo) incorporation, clonogenesis assays, cell cycle analysis and magnetic resonance imaging in living mice. Additionally, V3Nter switches off the β-catenin target gene expression signature in vivo. Moreover, experiments with β-catenin allele-targeted cells showed that the ΔS45 β-catenin allele hampers, but does not abrogate, inhibition of Wnt signaling by SFRP-1 or by the SFRP-like frizzled domain. Finally, neither SFRP-1 nor V3Nter affect β-catenin signaling in SW480 cells carrying nonfunctional Adenomatous polyposis coli. Thus, SFRP-1 and the SFRP-like molecule V3Nter can inhibit tumor growth of β-catenin-activated tumor cells in vivo.

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Abbreviations

C18:

collagen XVIII

CRC:

colorectal carcinoma

CRD:

cystein-rich domain

CRT:

β-catenin-T-cell factor-regulated transcription

FZC18:

frizzled module of collagen XVIII

GSEA:

gene set enrichment analysis

HEK293:

human embryonic kidney 293

Nter:

N-terminal

SFRP:

secreted frizzled-related protein

V2:

variant 2

V3:

variant 3

WT:

wild-type

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Acknowledgements

We thank technical assistance from C Rocher at the Small Animal Imaging Platform, A Fautrel and P Bellaud at the Core HistoPathology Platform, R Le Guével at the ImpactCell Platform, D Le Quilleuc and B Turlin at the Center for Biological Ressources and the animal husbandry group. We are indebted to B Vogelstein, S Bayling and H Suzuki, R Moon, R Nusse, T Pihlajaniemi and J Pouyssegur for generously providing cells, cDNAs or antibodies and to Christine Perret (Institut Cochin, Department Endocrinology, Metabolism and Cancer, Paris, France) for critical reading of the paper. This study was supported by Institut National de la Santé et de la Recherche Médicale, Institut National du Cancer, Agence Nationale de la Recherche (Emergence-BIO Program 2008), Université de Rennes 1, Région Bretagne.

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Authors and Affiliations

  1. INSERM, UMR991, Liver Metabolisms and Cancer, Rennes, France

    E Lavergne, I Hendaoui, C Coulouarn, C Ribault, J Leseur, S Mebarki, A Corlu, B Clément & O Musso

  2. Université de Rennes 1, Rennes, France

    E Lavergne, I Hendaoui, C Coulouarn, C Ribault, J Leseur, P-A Eliat, S Mebarki, A Corlu, B Clément & O Musso

  3. Centre Régional de Lutte contre le Cancer, Rennes, France

    J Leseur

  4. PRISM, IFR 140, Biogenouest, Rennes, France

    P-A Eliat

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Correspondence to O Musso.

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Competing interests

The following authors are coauthors of a patent owned by INSERM (Institut National de la Santé et de la Recherche Médicale) on the therapeutic use of the frizzled module of collagen XVIII: E Lavergne, I Hendaoui, J Leseur, B Clément and O Musso. C Coulouarn, C Ribault, P-A Eliat, S Mebarki and A Corlu declare no potential conflict of interest.

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Lavergne, E., Hendaoui, I., Coulouarn, C. et al. Blocking Wnt signaling by SFRP-like molecules inhibits in vivo cell proliferation and tumor growth in cells carrying active β-catenin. Oncogene 30, 423–433 (2011). https://doi.org/10.1038/onc.2010.432

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