The metastasis-associated gene S100A4 is a novel target of beta-catenin/T-cell factor signaling in colon cancer.

BACKGROUND & AIMS Activation of the Wnt/beta-catenin pathway is frequently observed in colorectal cancers. Our aim was to elucidate the impact of gain-of-function beta-catenin on the metastasis-associated gene S100A4 in human colon cancer cell lines and tumors. METHODS We analyzed cell lines heterozygous for gain-of-function and wild-type beta-catenin, and variants homozygous for gain- or loss-of-function mutation in beta-catenin, for S100A4 expression, cell motility, and in vivo metastasis. beta-catenin-mediated S100A4 promoter activation was tested by reporter assays. For human colon carcinomas, S100A4 expression, beta-catenin genotype, and metachronous metastasis were correlated. RESULTS We identified S100A4 as the most regulated gene by gain-of-function beta-catenin using a 10K microarray. Cell lines with gain-of-function beta-catenin expressed up to 60-fold elevated S100A4 levels, displayed strongly increased migration and invasion in vitro, and induced metastasis in mice. S100A4 small interfering RNA, beta-catenin small interfering RNA, or dominant negative T-cell factor (TCF) knocked down S100A4 and blocked biological effects. S100A4 complementary DNA transfection increased migration and invasion. We identified a TCF binding site within the S100A4 promoter and demonstrated the direct binding of heterodimeric beta-catenin/TCF complexes. Reporter assays confirmed the beta-catenin-induced S100A4 promoter activity. Furthermore, S100A4 mRNA expression was increased in primary colon cancers, which later developed distant metastases, compared to non-metastasizing tumors. Colon tumors heterozygous for gain-of-function beta-catenin showed concomitant nuclear beta-catenin localization, high S100A4 expression, and metastases. CONCLUSIONS S100A4 is a direct beta-catenin/TCF target, induces migration and invasion in vitro and metastasis in vivo, and has value for prognosis of metastasis formation in colon cancer patients.

[1]  David J Weber,et al.  S100A4, a Mediator of Metastasis* , 2006, Journal of Biological Chemistry.

[2]  D. Helfman,et al.  The metastasis associated protein S100A4: role in tumour progression and metastasis , 2005, British Journal of Cancer.

[3]  M. Grigorian,et al.  Suppression of tumor development and metastasis formation in mice lacking the S100A4(mts1) gene. , 2005, Cancer research.

[4]  C. Heizmann,et al.  S100 proteins in mouse and man: from evolution to function and pathology (including an update of the nomenclature). , 2004, Biochemical and biophysical research communications.

[5]  F. Brembeck,et al.  Essential role of BCL9-2 in the switch between beta-catenin's adhesive and transcriptional functions. , 2004, Genes & development.

[6]  K. Kinzler,et al.  Cancer genes and the pathways they control , 2004, Nature Medicine.

[7]  J. Chan,et al.  Prognostic and Diagnostic Significance of β-Catenin Nuclear Immunostaining in Colorectal Cancer , 2004, Clinical Cancer Research.

[8]  J. Minna,et al.  β-catenin inhibits cell growth of a malignant mesothelioma cell line, NCI-H28, with a 3p21.3 homozygous deletion , 2003, Oncogene.

[9]  Lin Li,et al.  Chromatin immunoprecipitation: a tool for studying histone acetylation and transcription factor binding. , 2003, Methods.

[10]  R. Gaynor,et al.  Functional Correlates of Mutations in β-Catenin Exon 3 Phosphorylation Sites* , 2003, Journal of Biological Chemistry.

[11]  T. Minamoto,et al.  β-catenin and ras oncogenes detect most human colorectal cancer , 2003 .

[12]  K. B. Pedersen,et al.  Nuclear localization of the metastasis‐related protein S100A4 correlates with tumour stage in colorectal cancer , 2003, The Journal of pathology.

[13]  S. Y. Park,et al.  Development and applications of a beta-catenin oligonucleotide microarray: beta-catenin mutations are dominantly found in the proximal colon cancers with microsatellite instability. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[14]  Hans Clevers,et al.  Caught up in a Wnt storm: Wnt signaling in cancer. , 2003, Biochimica et biophysica acta.

[15]  L. Siracusa,et al.  The Cohesin SMC3 Is a Target the for β-Catenin/TCF4 Transactivation Pathway* , 2003, Journal of Biological Chemistry.

[16]  H. Kreipe,et al.  Prognostic significance of calcium-binding protein S100A4 in colorectal cancer. , 2002, Gastroenterology.

[17]  Hans Clevers,et al.  The β-Catenin/TCF-4 Complex Imposes a Crypt Progenitor Phenotype on Colorectal Cancer Cells , 2002, Cell.

[18]  P. Hohenberger,et al.  Expression of multidrug resistance genes MVP, MDR1, and MRP1 determined sequentially before, during, and after hyperthermic isolated limb perfusion of soft tissue sarcoma and melanoma patients. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  Baljit Singh,et al.  Oncogenic β-Catenin Is Required for Bone Morphogenetic Protein 4 Expression in Human Cancer Cells , 2002 .

[20]  Xi He,et al.  Control of β-Catenin Phosphorylation/Degradation by a Dual-Kinase Mechanism , 2002, Cell.

[21]  M. Grigorian,et al.  Liprin β1, a Member of the Family of LAR Transmembrane Tyrosine Phosphatase-interacting Proteins, Is a New Target for the Metastasis-associated Protein S100A4 (Mts1)* , 2002, The Journal of Biological Chemistry.

[22]  P. Rudland,et al.  S100A4 (p9Ka) protein in colon carcinoma and liver metastases: association with carcinoma cells and T-lymphocytes , 2002, British Journal of Cancer.

[23]  R. Knuechel,et al.  Variable β-catenin expression in colorectal cancers indicates tumor progression driven by the tumor environment , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[24]  P. Schlag,et al.  Hyperthermia-induced Nuclear Translocation of Transcription Factor YB-1 Leads to Enhanced Expression of Multidrug Resistance-related ABC Transporters* , 2001, The Journal of Biological Chemistry.

[25]  J. Minna,et al.  Genetic alteration of the β-catenin gene (CTNNB1) in human lung cancer and malignant mesothelioma and identification of a new 3p21.3 homozygous deletion , 2001, Oncogene.

[26]  S. Maeda,et al.  The role of S100A4 gene encoding an S100-related calcium-binding protein in human bile duct adenocarcinoma cell lines: correlation of S100A4 expression and invasive growth in Matrigel Matrix. , 2000, International Journal of Molecular Medicine.

[27]  H. Clevers,et al.  Linking Colorectal Cancer to Wnt Signaling , 2000, Cell.

[28]  P. Polakis.  Wnt signaling and cancer. , 2000, Genes & development.

[29]  O. T. Odegaard,et al.  S100A4 involvement in metastasis: deregulation of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in osteosarcoma cells transfected with an anti-S100A4 ribozyme. , 1999, Cancer research.

[30]  Paul Polakis,et al.  The metalloproteinase matrilysin is a target of β-catenin transactivation in intestinal tumors , 1999, Oncogene.

[31]  Walter Birchmeier,et al.  Hot papers in cell biology - J. Behrens, J.P. von Kries, M. Kuehl, L. Bruhn, D. Wedlich, R. Grosschedl, W. Birchmeier: "Functional interaction of beta-catenin with the transcription factor LEF-1" - Comments by Walter Birchmeier , 1999 .

[32]  Frank McCormick,et al.  β-Catenin regulates expression of cyclin D1 in colon carcinoma cells , 1999, Nature.

[33]  W. Bodmer,et al.  Genetic pathways in colorectal and other cancers. , 1999, European journal of cancer.

[34]  Paul Polakis,et al.  The oncogenic activation of β-catenin , 1999 .

[35]  R. Barraclough.  Calcium-binding protein S100A4 in health and disease. , 1998, Biochimica et biophysica acta.

[36]  Hans Clevers,et al.  The Xenopus Wnt effector XTcf-3 interacts with Groucho-related transcriptional repressors , 1998, Nature.

[37]  B. Geiger,et al.  Differential molecular interactions of beta-catenin and plakoglobin in adhesion, signaling and cancer. , 1998, Current opinion in cell biology.

[38]  W. Birchmeier,et al.  Functional interaction of an axin homolog, conductin, with beta-catenin, APC, and GSK3beta. , 1998, Science.

[39]  Bert Vogelstein,et al.  Mutational Analysis of the APC/β-Catenin/Tcf Pathway in Colorectal Cancer , 1998 .

[40]  Akira Kikuchi,et al.  Axin, a negative regulator of the Wnt signaling pathway, forms a complex with GSK‐3β and β‐catenin and promotes GSK‐3β‐dependent phosphorylation of β‐catenin , 1998 .

[41]  K. Wada,et al.  Increased expression of S100A4, a metastasis-associated gene, in human colorectal adenocarcinomas. , 1997, Clinical cancer research : an official journal of the American Association for Cancer Research.

[42]  Stephen W. Byers,et al.  Serine Phosphorylation-regulated Ubiquitination and Degradation of β-Catenin* , 1997, The Journal of Biological Chemistry.

[43]  W F Bodmer,et al.  Beta-catenin mutations in cell lines established from human colorectal cancers. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Jörg Stappert,et al.  β‐catenin is a target for the ubiquitin–proteasome pathway , 1997 .

[45]  Paul Polakis,et al.  Stabilization of β-Catenin by Genetic Defects in Melanoma Cell Lines , 1997, Science.

[46]  Hans Clevers,et al.  Activation of β-Catenin-Tcf Signaling in Colon Cancer by Mutations in β-Catenin or APC , 1997, Science.

[47]  Yohko Nakamura,et al.  Expression of antisense RNA to S100A4 gene encoding an S100-related calcium-binding protein suppresses metastatic potential of high-metastatic Lewis lung carcinoma cells , 1997, Oncogene.

[48]  M. Grigorian,et al.  Metastasis of mammary carcinomas in GRS/A hybrid mice transgenic for the mts1 gene. , 1996, Oncogene.

[49]  P. Rudland,et al.  Expression of the calcium-binding protein S100A4 (p9Ka) in MMTV-neu transgenic mice induces metastasis of mammary tumours. , 1996, Oncogene.

[50]  Michael Kühl,et al.  Functional interaction of β-catenin with the transcription factor LEF-1 , 1996, Nature.

[51]  A. Debant,et al.  The multidomain protein Trio binds the LAR transmembrane tyrosine phosphatase, contains a protein kinase domain, and has separate rac-specific and rho-specific guanine nucleotide exchange factor domains. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[52]  E. Hay,et al.  Transformations between epithelium and mesenchyme: normal, pathological, and experimentally induced. , 1995, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[53]  M. Grigorian,et al.  The mts1 gene and control of tumor metastasis. , 1993, Gene.

[54]  C. Heizmann,et al.  S100 alpha, CAPL, and CACY: molecular cloning and expression analysis of three calcium-binding proteins from human heart. , 1992, Biochemistry.

[55]  B. Vogelstein,et al.  A genetic model for colorectal tumorigenesis , 1990, Cell.

[56]  R. Gilbertson,et al.  ERBB2 up-regulates S100A4 and several other prometastatic genes in medulloblastoma. , 2003, Cancer research.

[57]  L. Mazzucchelli.  Protein S100A4: too long overlooked by pathologists? , 2002, The American journal of pathology.