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Opposing roles of netrin-1 and the dependence receptor DCC in cancer cell invasion, tumor growth and metastasis

Oncogene volume 26, pages 5615–5625 (2007)Cite this article

Abstract

Deleted in colon cancer (DCC) and UNC5 function as netrin dependence receptors by inducing apoptosis in the absence of their ligand and accordingly were recently designated as putative conditional tumor suppressors. Herein, we determined whether netrin-1 and its receptors are implicated in cancer cell invasion and tumor progression. Expression of DCC, UNC5 and adenosine A2B-receptors (A2B-Rs) was investigated by reverse transcription polymerase chain reaction in human colon cancer cells. The impact of DCC restitution and netrin-1 was evaluated on collagen type I invasion, tumor growth and metastasis in nude mice, cancer cell survival and gene expression profiling. Flow cytometry, poly(ADP-ribose)polymerase-1 and caspase-8 activation were used to evaluate the impact of DCC on cell death. Both netrin-1 and A2B-R activation induced the invasive phenotype through the Rho–Rho kinase axis in DCC-deficient human colorectal cancer cells. Restitution of wild-type DCC blocked invasion induced by netrin-1, A2B-R agonist and other agents. Ectopic expression of netrin-1 led to increased growth of human colon tumor xenografts in athymic mice. Conversely, introduction of wt-DCC in kidney MDCKts.src-ggl cells strongly inhibited metastasis in lymph nodes and lungs and increased sensitivity to apoptosis in hypoxia. DNA microarrays revealed that netrin and DCC had common and divergent impacts on gene expression linked to cell cycle, survival, surface signaling and adhesion. Our findings underscore that netrin is a potent invasion and tumor growth-promoting agent and that DCC is a metastasis suppressor gene targeting both proinvasive and survival pathways in a cumulative manner.

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Abbreviations

A2B-R:

adenosine A2B-receptor

DCC:

deleted in colon cancer

EGF-R:

epidermal growth factor receptor

GCOS:

GeneChip Operating System v1.4

GPCR:

G-protein-coupled receptors

PARP:

poly(ADP-ribose)polymerase-1

PI3K:

phosphatidylinositol 3-kinase

PKA:

protein kinase A

PLCβ:

phospholipase Cβ

ROK:

Rho-kinase

RT–PCR:

reverse transcription–polymerase chain reaction

TGFα:

transforming growth factor α

TFFs:

trefoil factors

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Acknowledgements

This work is supported by INSERM, ARC, the Fund for Scientific Research Flanders (Brussels, Belgium) and the Scientific Exchange Program between the Flemish community and France (Grant I.2006/2007.03). We are indebted to Dr Lamarche-Vane and to Dr Mehlen for providing the initial batch of recombinant netrin-1 and the expression vectors encoding netrin-1 and the DCC.

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Authors and Affiliations

  1. INSERM, U673, Paris, France

    S Rodrigues, O De Wever & C Gespach

  2. Université Pierre et Marie Curie-Paris 6, Faculté de Médecine, Laboratory of Molecular and Clinical Oncology of Solid Tumors, Paris, France

    S Rodrigues & C Gespach

  3. Ghent University Hospital, Laboratory of Experimental Cancerology, Ghent, Belgium

    O De Wever & E Bruyneel

  4. Genome Explorations Inc., Memphis, TN, USA

    R J Rooney

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  1. S Rodrigues
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  2. O De Wever
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  3. E Bruyneel
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  5. C Gespach
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Correspondence to C Gespach.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Rodrigues, S., De Wever, O., Bruyneel, E. et al. Opposing roles of netrin-1 and the dependence receptor DCC in cancer cell invasion, tumor growth and metastasis. Oncogene 26, 5615–5625 (2007). https://doi.org/10.1038/sj.onc.1210347

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