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MLK3 is critical for breast cancer cell migration and promotes a malignant phenotype in mammary epithelial cells

Oncogene volume 29, pages 4399–4411 (2010)Cite this article

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Abstract

The malignant phenotype in breast cancer is driven by aberrant signal transduction pathways. Mixed-lineage kinase-3 (MLK3) is a mammalian mitogen-activated protein kinase kinase kinase (MAP3K) that activates multiple MAPK pathways. Depending on the cellular context, MLK3 has been implicated in apoptosis, proliferation, migration and differentiation. Here we investigated the effect of MLK3 and its signaling to MAPKs in the acquisition of malignancy in breast cancer. We show that MLK3 is highly expressed in breast cancer cells. We provide evidence that MLK3's catalytic activity and signaling to c-jun N-terminal kinase (JNK) is required for migration of highly invasive breast cancer cells and for MLK3-induced migration of mammary epithelial cells. Expression of active MLK3 is sufficient to induce the invasion of mammary epithelial cells, which requires AP-1 activity and is accompanied by the expression of several proteins corresponding to AP-1-regulated invasion genes. To assess MLK3's contribution to the breast cancer malignant phenotype in a more physiological setting, we implemented a strategy to inducibly express active MLK3 in the preformed acini of MCF10A cells grown in 3D Matrigel. Induction of MLK3 expression dramatically increases acinar size and modestly perturbs apicobasal polarity. Remarkably, MLK3 expression induces luminal repopulation and suppresses the expression of the pro-apoptotic protein BimEL, as has been observed in Her2/Neu-expressing acini. Taken together, our data show that MLK3–JNK–AP-1 signaling is critical for breast cancer cell migration and invasion. Our current study uncovers both a proliferative and novel antiapoptotic role for MLK3 in the acquisition of a malignant phenotype in mammary epithelial cells. Thus, MLK3 may be an important therapeutic target for the treatment of invasive breast cancer.

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Acknowledgements

This work was supported by an MSU-Health and Biomedical Research Institute seed grant (KAG HBRI-729) and a DoD Breast Cancer Idea Award (KAG BC085020). Confocal microscopes were provided through the MSU Center for Advanced Microscopy. We are grateful to Cephalon Inc. for CEP-11004; Ariad Pharmaceuticals for AP21967 and for parental inducible expression vectors; Richard Mulligan (Harvard Medical School) for the 293GPG packaging cell line and Daniel DiMaio (Yale Cancer Center) for the pBabe-Tam67 construct. We thank Mauricio Reginato (Drexel University) for advice on Bim immunoblotting and Bradley Smith (Department of Physiology, MSU) for help with the morphometric analysis of the MCF10A cells grown in 3D culture. We appreciate the constructive input from the members of the MSU-Breast Cancer Signaling Networks Consortium.

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  1. Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, USA

    J Chen

  2. Department of Physiology, Michigan State University, East Lansing, MI, USA

    E M Miller & K A Gallo

  3. Cell and Molecular Biology Program, Michigan State University, East Lansing, MI, USA

    K A Gallo

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Correspondence to K A Gallo.

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Chen, J., Miller, E. & Gallo, K. MLK3 is critical for breast cancer cell migration and promotes a malignant phenotype in mammary epithelial cells. Oncogene 29, 4399–4411 (2010). https://doi.org/10.1038/onc.2010.198

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