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SPON2, a newly identified target gene of MACC1, drives colorectal cancer metastasis in mice and is prognostic for colorectal cancer patient survival

Oncogene volume 35, pages 5942–5952 (2016)Cite this article

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Abstract

MACC1 (metastasis associated in colon cancer 1) is a prognostic biomarker for tumor progression, metastasis and survival of a variety of solid cancers including colorectal cancer (CRC). Here we aimed to identify the MACC1-induced transcriptome and key players mediating the MACC1-induced effects in CRC. We performed microarray analyses using CRC cells ectopically overexpressing MACC1. We identified more than 1300 genes at least twofold differentially expressed, including the gene SPON2 (Spondin 2) as 90-fold upregulated transcriptional target of MACC1. MACC1-dependent SPON2 expression regulation was validated on mRNA and protein levels in MACC1 high (endogenously or ectopically) and low (endogenously or by knockdown) expressing cells. Chromatin immunoprecipitation analysis demonstrated the binding of MACC1 to the gene promoter of SPON2. In cell culture, ectopic SPON2 overexpression induced cell viability, migration, invasion and colony formation in endogenously MACC1 and SPON2 low expressing cells, whereas SPON2 knockdown reduced proliferative, migratory and invasive abilities in CRC cells with high endogenous MACC1 and SPON2 expression. In intrasplenically transplanted NOD/SCID mice, metastasis induction was analyzed with control or SPON2-overexpressing CRC cells. Tumors with SPON2 overexpression induced liver metastasis (vs control animals without any metastases, P=0.0036). In CRC patients, SPON2 expression was determined in primary tumors (stages I–III), and survival time was analyzed by Kaplan–Meier method. CRC patients with high SPON2 expressing primary tumors demonstrated 8 months shorter metastasis-free survival (MFS) compared with patients with low SPON2 levels (P=0.053). Combining high levels of SPON2 and MACC1 improved the identification of high-risk patients with a 20-month shorter MFS vs patients with low biomarker expression. In summary, SPON2 is a transcriptional target of the metastasis gene MACC1. SPON2 induces cell motility in vitro and CRC metastasis in mice. In patients, SPON2 serves as prognostic indicator for CRC metastasis and survival, and might represent a promising target for therapeutic approaches.

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Acknowledgements

We are thankful to Wolfgang Kemmner and his laboratory for the help with the microarray experiments. We thank Maria Stecklum and Nadine Bäsler for support with the in vivo bioluminescence imaging, and Jutta Aumann for assistance on expression analyses. We thank Manisha Juneja and Katharina Ilm for help with the combinatorial biomarker analysis in the CRC patients. We are grateful to Markus Niederstrasser for statistical advice. This work was supported by the German Cancer Consortium (DKTK).

Author contributions

FS and US conceived and designed the experiments and wrote the manuscript. FS and JS performed the in vitro experiments. FS, QW, MRH and MAA-N analyzed the microarray data. FS, ML and IF designed and performed the in vivo studies. MD, DK and WW evaluated the experimental data and contributed to the revised version of the manuscript. PMS evaluated the clinical data. All authors have read and approved the final manuscript.

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Authors and Affiliations

  1. Experimental and Clinical Research Center, Charité Universitätsmedizin Berlin and Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany

    F Schmid, Q Wang, M Dahlmann, D Kobelt, W Walther, J Smith & U Stein

  2. Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany

    M R Huska, M A Andrade-Navarro, M Lemm & I Fichtner

  3. Charité Comprehensive Cancer Center, Berlin, Germany

    P M Schlag

  4. German Cancer Consortium (DKTK), Heidelberg, Germany

    U Stein

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  1. F Schmid
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Correspondence to U Stein.

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Schmid, F., Wang, Q., Huska, M. et al. SPON2, a newly identified target gene of MACC1, drives colorectal cancer metastasis in mice and is prognostic for colorectal cancer patient survival. Oncogene 35, 5942–5952 (2016). https://doi.org/10.1038/onc.2015.451

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