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mTORC1 upregulation via ERK-dependent gene expression change confers intrinsic resistance to MEK inhibitors in oncogenic KRas-mutant cancer cells

Oncogene volume 34, pages 5607–5616 (2015)Cite this article

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Abstract

Cancer cells harboring oncogenic BRaf mutants, but not oncogenic KRas mutants, are sensitive to MEK inhibitors (MEKi). The mechanism underlying the intrinsic resistance to MEKi in KRas-mutant cells is under intensive investigation. Here, we pursued this mechanism by live imaging of extracellular signal-regulated kinases (ERK) and mammalian target of rapamycin complex 1 (mTORC1) activities in oncogenic KRas or BRaf-mutant cancer cells. We established eight cancer cell lines expressing Förster resonance energy transfer (FRET) biosensors for ERK activity and S6K activity, which was used as a surrogate marker for mTORC1 activity. Under increasing concentrations of MEKi, ERK activity correlated linearly with the cell growth rate in BRaf-mutant cancer cells, but not KRas-mutant cancer cells. The administration of PI3K inhibitors resulted in a linear correlation between ERK activity and cell growth rate in KRas-mutant cancer cells. Intriguingly, mTORC1 activity was correlated linearly with the cell growth rate in both BRaf-mutant cancer cells and KRas-mutant cancer cells. These observations suggested that mTORC1 activity had a pivotal role in cell growth and that the mTORC1 activity was maintained primarily by the ERK pathway in BRaf-mutant cancer cells and by both the ERK and PI3K pathways in KRas-mutant cancer cells. FRET imaging revealed that MEKi inhibited mTORC1 activity with slow kinetics, implying transcriptional control of mTORC1 activity by ERK. In agreement with this observation, MEKi induced the expression of negative regulators of mTORC1, including TSC1, TSC2 and Deptor, which occurred more significantly in BRaf-mutant cells than in KRas-mutant cells. These findings suggested that the suppression of mTORC1 activity and induction of negative regulators of mTORC1 in cancer cells treated for at least 1 day could be used as surrogate markers for the MEKi sensitivity of cancer cells.

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Acknowledgements

We are grateful to the members of the Matsuda Laboratory for their helpful input. Y Inaoka, K Hirano, A Katsumata, N Nishimoto, N Nonaka and A Kawagishi are also to be thanked for their technical assistance. KA and MM were supported by the Research Program of Innovative Cell Biology by Innovative Technology (the ‘Cell Innovation’ Program) and Platform for Dynamic Approaches to Living System from the Ministry of Education, Culture, Sports and Science, Japan. KA was supported by a Grant-in-Aid for Young Scientists (B) (23701052) and a Grant-in-Aid for Scientific Research (B). NK and YF were supported by a Grant-in-Aid for JSPS Fellows.

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Authors and Affiliations

  1. Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto, Japan,

    N Komatsu & M Matsuda

  2. Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Kyoto, Japan,

    Y Fujita & M Matsuda

  3. Imaging Platform for Spatio-Temporal Information, Graduate School of Medicine, Kyoto University, Kyoto, Japan,

    K Aoki

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  1. N Komatsu
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Correspondence to K Aoki.

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Komatsu, N., Fujita, Y., Matsuda, M. et al. mTORC1 upregulation via ERK-dependent gene expression change confers intrinsic resistance to MEK inhibitors in oncogenic KRas-mutant cancer cells. Oncogene 34, 5607–5616 (2015). https://doi.org/10.1038/onc.2015.16

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