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miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis

Nature Cell Biology volume 12, pages 247–256 (2010)Cite this article

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Abstract

MicroRNAs (miRNAs) are increasingly implicated in regulating the malignant progression of cancer. Here we show that miR-9, which is upregulated in breast cancer cells, directly targets CDH1, the E-cadherin-encoding messenger RNA, leading to increased cell motility and invasiveness. miR-9-mediated E-cadherin downregulation results in the activation of β-catenin signalling, which contributes to upregulated expression of the gene encoding vascular endothelial growth factor (VEGF); this leads, in turn, to increased tumour angiogenesis. Overexpression of miR-9 in otherwise non-metastatic breast tumour cells enables these cells to form pulmonary micrometastases in mice. Conversely, inhibiting miR-9 by using a 'miRNA sponge' in highly malignant cells inhibits metastasis formation. Expression of miR-9 is activated by MYC and MYCN, both of which directly bind to the mir-9-3 locus. Significantly, in human cancers, miR-9 levels correlate with MYCN amplification, tumour grade and metastatic status. These findings uncover a regulatory and signalling pathway involving a metastasis-promoting miRNA that is predicted to directly target expression of the key metastasis-suppressing protein E-cadherin.

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Figure 1: miR-9 directly targets CDH1 and increases cell motility and invasiveness.
Figure 2: miR-9 increases VEGF levels in an E-cadherin-dependent and β-catenin-dependent manner.
Figure 3: miR-9 induces angiogenesis, mesenchymal marker expression and metastasis of the SUM149 epithelial tumours.
Figure 4: Inhibiting miR-9 suppresses metastasis.
Figure 5: miR-9 expression is activated by MYC/MYCN.
Figure 6: miR-9 levels correlate with MYCN amplification and metastatic status in human cancers.

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Acknowledgements

We thank Johannes Schulte, Martin Eilers, Rosa Noguera, Margaret Ebert and Phillip Sharp for providing tumour samples and reagents; the Histology Core Lab at Massachusetts Institute of Technology (MIT) and Memorial Sloan-Kettering Cancer Center (MSKCC) for assistance with sectioning and immunohistochemistry; and members of the Weinberg laboratory for useful discussions. L.M. is a recipient of a Life Sciences Research Foundation Fellowship, a Margaret and Herman Sokol Award, and a National Institutes of Health (NIH) Pathway to Independence Award (K99/R00). J.Y. and E.P. are supported by a Howard Hughes Medical Institute Undergraduate Fellowship. J.T.-F. is supported by the MSKCC Cancer Core Grant. T.T.O. and S.V. are recipients of a US Department of Defense Breast Cancer Research Program Predoctoral Fellowship. R.A.W. is an American Cancer Society Research Professor and a Daniel K. Ludwig Cancer Research Professor. This research is supported by a NIH grant to R.A.W. and the Ludwig Center for Molecular Oncology at MIT.

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Author notes

  1. Jennifer Young and Harsha Prabhala: These authors contributed equally to this work.

Authors and Affiliations

  1. Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, 02142, Massachusetts, USA

    Li Ma, Jennifer Young, Elizabeth Pan, Ferenc Reinhardt, Tamer T. Onder, Scott Valastyan & Robert A. Weinberg

  2. Massachusetts Institute of Technology Ludwig Center for Molecular Oncology, Cambridge, 02142, Massachusetts, USA

    Li Ma, Tamer T. Onder, Scott Valastyan & Robert A. Weinberg

  3. Medical Scientist Training Program, University of Virginia, Charlottesville, 22908, Virginia, USA

    Harsha Prabhala

  4. Center for Medical Genetics, Ghent University Hospital, Ghent, 9000, Belgium

    Pieter Mestdagh, Frank Speleman & Jo Vandesompele

  5. Department of Tumor Genetics, German Cancer Center, Im Neuenheimer Feld 280, Heidelberg, 69120, Germany

    Daniel Muth & Frank Westermann

  6. Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, 10021, New York, USA

    Julie Teruya-Feldstein

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Contributions

L.M. conceived the project. R.A.W. supervised research. L.M. and H.P. designed experiments. L.M., J.Y., H.P., E.P., J.T.-F. and F.R. performed most of the experiments and analysed data. P.M., D.M., F.W., F.P. and J.V. contributed MYCN and ChIP-on-chip data. T.T.O. contributed some of the constructs and shared unpublished observations. S.V. modified the miRNA sponge design for stable expression. L.M. and R.A.W. wrote the manuscript.

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Correspondence to Robert A. Weinberg.

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Ma, L., Young, J., Prabhala, H. et al. miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nat Cell Biol 12, 247–256 (2010). https://doi.org/10.1038/ncb2024

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