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Epigenetic inactivation of SFRP genes allows constitutive WNT signaling in colorectal cancer

Nature Genetics volume 36, pages 417–422 (2004)Cite this article

Abstract

Aberrant WNT pathway signaling is an early progression event in 90% of colorectal cancers1. It occurs through mutations mainly of APC and less often of CTNNB1 (encoding β-catenin)1,2,3 or AXIN2 (encoding axin-2, also known as conductin)4. These mutations allow ligand-independent WNT signaling that culminates in abnormal accumulation of free β-catenin in the nucleus1,2,3. We previously identified frequent promoter hypermethylation and gene silencing of the genes encoding secreted frizzled-related proteins (SFRPs) in colorectal cancer5. SFRPs possess a domain similar to one in the WNT-receptor frizzled proteins and can inhibit WNT receptor binding to downregulate pathway signaling during development6,7,8,9,10. Here we show that restoration of SFRP function in colorectal cancer cells attenuates WNT signaling even in the presence of downstream mutations. We also show that the epigenetic loss of SFRP function occurs early in colorectal cancer progression and may thus provide constitutive WNT signaling that is required to complement downstream mutations in the evolution of colorectal cancer.

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Figure 1: Colorectal cancer cells express various WNTs and the WNT signaling pathway can be suppressed by SFRPs.
Figure 2: Demethylation and re-expression of SFRP genes and suppression of the WNT pathway in DKO cells.
Figure 3: Upregulation of the canonical WNT pathway by WNT1, β-catenin and LEF1, which can be suppressed by SFRPs in colon cancer cells.
Figure 4: SFRPs suppress β-catenin and CRT target gene expression and induce apoptosis.
Figure 5: Methylation and expression analysis of SFRP genes in primary colorectal cancers (a) Bisulfite sequencing analysis of SFRP1 in peripheral blood lymphocytes (PBLs), normal colon tissue, and paired normal counterparts (N) and tumor tissues (T) from three individuals with colorectal cancer.
Figure 6: Methylation analysis of SFRP genes in ACF and colorectal adenoma samples.

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Acknowledgements

We thank B. Vogelstein for providing pGL3-OT and pGL3-OF reporters and for comments on the manuscript; S. Eguchi, B. Reese, K. Ogi and K. Akino for technical advice; and K. Bender for technical support. S.D.M. is an Investigator in the Howard Hughes Medical Institute and supported by the National Cancer Institute. T.P.P. is supported by grants from the US Public Health Service and the National Cancer Institute. This work was supported by a grant from the National Institute of Environmental Health Services.

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Authors and Affiliations

  1. Division of Tumor Biology, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins Medical Institutions, 1650 Orleans Street, Baltimore, 21231, Maryland, USA

    Hiromu Suzuki, D Neil Watkins, Kam-Wing Jair, Kornel E Schuebel, Bin Yang, Yoshimitsu Akiyama, James G Herman & Stephen B Baylin

  2. First Department of Internal Medicine, Sapporo Medical University, Sapporo, Japan

    Hiromu Suzuki, Minoru Toyota & Kohzoh Imai

  3. Howard Hughes Medical Institute, Cleveland, 44106, Ohio, USA

    Sanford D Markowitz

  4. Department of Medicine and Ireland Cancer Center, Case Western Reserve University and University Hospitals of Cleveland, Cleveland, 44106, Ohio, USA

    Sanford D Markowitz & Wei Dong Chen

  5. Department of Pathology, Case Western Reserve University, Cleveland, 44106, Ohio, USA

    Theresa P Pretlow

  6. Department of Pathology, University of Maastricht, Maastricht, 6200 MD, The Netherlands

    Manon van Engeland

  7. Department of Molecular Biology, Cancer Research Institute, Sapporo Medical University, Sapporo, Japan

    Minoru Toyota & Takashi Tokino

  8. PRESTO, JST, Kawaguchi, Japan

    Minoru Toyota

  9. Department of Clinical Laboratory Science, Yamaguchi University School of Medicine, Yamaguchi, Japan

    Yuji Hinoda

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Correspondence to Stephen B Baylin.

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Competing interests

The commercial rights to the MSP technique belong to Oncomethylome. S.B.B. and J.G.H. serve as consultants to Oncomethylome and are entitled to royalties from any commercial use of this procedure.

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Suzuki, H., Watkins, D., Jair, KW. et al. Epigenetic inactivation of SFRP genes allows constitutive WNT signaling in colorectal cancer. Nat Genet 36, 417–422 (2004). https://doi.org/10.1038/ng1330

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