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Defining MAP3 kinases required for MDA-MB-231 cell tumor growth and metastasis

Oncogene volume 31, pages 3889–3900 (2012)Cite this article

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Abstract

Analysis of patient tumors suggests that multiple MAP3 kinases (MAP3Ks) are critical for growth and metastasis of cancer cells. MAP3Ks selectively control the activation of extracellular signal-regulated kinase 1/2 (ERK1/2), Jun N-terminal kinase (JNK), p38 and ERK5 in response to receptor tyrosine kinases and GTPases. We used MDA-MB-231 cells because of their ability to metastasize from the breast fat pad to distant lymph nodes for an orthotopic xenograft model to screen the function of seven MAP3Ks in controlling tumor growth and metastasis. Stable short hairpin RNA (shRNA) knockdown was used to inhibit the expression of each of the seven MAP3Ks, which were selected for their differential regulation of the MAPK network. The screen identified two MAP3Ks, MEKK2 and MLK3, whose shRNA knockdown caused significant inhibition of both tumor growth and metastasis. Neither MEKK2 nor MLK3 have been previously shown to regulate tumor growth and metastasis in vivo. These results demonstrated that MAP3Ks, which differentially activate JNK, p38 and ERK5, are necessary for xenograft tumor growth and metastasis of MDA-MB-231 tumors. The requirement for MAP3Ks signaling through multiple MAPK pathways explains why several members of the MAPK network are activated in cancer. MEKK2 was required for epidermal growth factor receptor and Her2/Neu activation of ERK5, with ERK5 being required for metastasis. Loss of MLK3 expression increased mitotic infidelity and apoptosis in vitro. Knockdown of MEKK2 and MLK3 resulted in increased apoptosis in orthotopic xenografts relative to control tumors in mice, inhibiting both tumor growth and metastasis; MEKK2 and MLK3 represent untargeted kinases in tumor biology for potential therapeutic development.

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Acknowledgements

This work was supported by NIH Grants GM30324 (GLJ), DK37871 (GLJ), GM059167 (HGD), U54CA156735 (GLJ and JES) and CA120881 (BC) and the University of North Carolina Cancer Research Fund (GLJ). We acknowledge the RNAi Screening Core and Lentiviral shRNA Core for support of this research.

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Authors and Affiliations

  1. Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    M R Cronan, K Nakamura, N L Johnson, D A Granger & G L Johnson

  2. Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    M R Cronan & H G Dohlman

  3. Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    M R Cronan, K Nakamura, N L Johnson, D A Granger, J-G Wang, N Mackman, H G Dohlman & G L Johnson

  4. Department of Molecular Pharmacology and Therapeutics, Stritch School of Medicine, Loyola University Chicago, Maywood, IL, USA

    B D Cuevas

  5. Division of Hematology and Oncology, Department of Medicine, McAllister Heart Institute, University of North Carolina School of Medicine, Chapel Hill, NC, USA

    J-G Wang & N Mackman

  6. Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, Durham, NC, USA

    J E Scott

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  1. M R Cronan
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Correspondence to G L Johnson.

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Cronan, M., Nakamura, K., Johnson, N. et al. Defining MAP3 kinases required for MDA-MB-231 cell tumor growth and metastasis. Oncogene 31, 3889–3900 (2012). https://doi.org/10.1038/onc.2011.544

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