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miR-145-dependent targeting of Junctional Adhesion Molecule A and modulation of fascin expression are associated with reduced breast cancer cell motility and invasiveness

Oncogene volume 29, pages 6569–6580 (2010)Cite this article

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Abstract

Micro RNAs are small non-coding RNAs, which regulate fundamental cellular and developmental processes at the transcriptional and translational level. In breast cancer, miR-145 expression is downregulated compared with healthy control tissue. As several predicted targets of miR-145 potentially regulate cell motility, we aimed at investigating a potential role for miR-145 in breast cancer cell motility and invasiveness. Assisted by Affymetrix array technology, we demonstrate that overexpression of miR-145 in MDA-MB-231, MCF-7, MDA-MB-468 and SK-BR-3 breast cancer cells and in Ishikawa endometrial carcinoma cells leads to a downregulation of the cell–cell adhesion protein JAM-A and of the actin bundling protein fascin. Moreover, podocalyxin and Serpin E1 mRNA levels were downregulated, and gamma-actin, transgelin and MYL9 were upregulated upon miR-145 overexpression. These miR-145-dependent expression changes drastically decreased cancer cell motility, as revealed by time-lapse video microscopy, scratch wound closure assays and matrigel invasion assays. Immunofluorescence microscopy demonstrated restructuring of the actin cytoskeleton and a change in cell morphology by miR-145 overexpression, resulting in a more cortical actin distribution, and reduced actin stress fiber and filopodia formation. Nuclear rotation was observed in 10% of the pre-miR-145 transfected MDA-MB-231 cells, accompanied by a reduction of perinuclear actin. Luciferase activation assays confirmed direct miR-145-dependent regulation of the 3′UTR of JAM-A, whereas siRNA-mediated knockdown of JAM-A expression resulted in decreased motility and invasiveness of MDA-MB-231 and MCF-7 breast cancer cells. Our data identify JAM-A and fascin as novel targets of miR-145, firmly establishing a role for miR-145 in modulating breast cancer cell motility. Our data provide a rationale for future miR-145-targeted approaches of antimetastatic cancer therapy.

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Acknowledgements

We would like to thank Birgit Pers, Ruth Goez, and Barbara Kloke for expert technical assistance, Professor Anna Starzinski-Powitz for providing Ishikawa cells, and Christian Adams MSc, for help with luciferase assays. Funding was provided by Grant NMRC/1023/2005 from the National Medical Research Council, Singapore (to GWY), by German Academic Exchange Service DAAD Grants A/08/15601 (to MV), and A/06/90277 (to SAI), and by the Förderkreis Universität Münster (to MG).

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  1. M Götte and C Mohr: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Gynecology and Obstetrics, Münster University Hospital, Münster, Germany

    M Götte, C Mohr, A-K Vaske, M Viola, S A Ibrahim & L Kiesel

  2. Department of Anatomy, National University of Singapore, Singapore

    C-Y Koo, Y H-F Teng, J-Y Low & G W Yip

  3. Institute of Physiology II, Münster University Hospital, Münster, Germany

    C Stock

  4. Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt

    S A Ibrahim

  5. Cell Adhesion and Cell Polarity Group, Institute of Medical Biochemistry, ZMBE, University of Münster, Münster, Germany

    S Peddibhotla & K Ebnet

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  1. M Götte
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Correspondence to M Götte or G W Yip.

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Götte, M., Mohr, C., Koo, CY. et al. miR-145-dependent targeting of Junctional Adhesion Molecule A and modulation of fascin expression are associated with reduced breast cancer cell motility and invasiveness. Oncogene 29, 6569–6580 (2010). https://doi.org/10.1038/onc.2010.386

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