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miR-141 regulates KEAP1 and modulates cisplatin sensitivity in ovarian cancer cells

Oncogene volume 32, pages 4284–4293 (2013)Cite this article

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Abstract

Epithelial ovarian cancer is the most lethal gynecological malignancy in the Western world. A major impediment for the successful treatment is the development of drug resistance. The molecular processes that contribute to resistance have been extensively studied; however, there is not much known about regulation by microRNAs (miRNAs). We compared miRNA expression profiles of an isogenic cisplatin-sensitive and -resistant ovarian cancer cell line pair (A2780/A2780 DDP) and found 27 miRNAs to be differentially expressed (⩾2-fold). Five of these, including the family members miR-141/200c, showed a correlation with cisplatin sensitivity in the NCI-60 panel. Overexpression of miR-141 resulted in enhanced resistance to cisplatin in ovarian cancer cell lines. We next correlated the expression level of miR-141 in 132 primary ovarian tumors (108 serous and 24 non-serous) with response to platinum-based chemotherapy. Although no differences were observed in the serous tumors, miR-141 levels were higher in non-serous ovarian tumors that did not respond well to therapy (platinum-free interval <6 months). We demonstrate that miR-141 directly targets KEAP1, and that downregulation of KEAP1 induces cisplatin resistance. Conversely, overexpression of KEAP1 significantly enhanced cisplatin sensitivity. Expression of KEAP1 with its 3′-UTR, and a 3′-UTR in which the miR-141 target site has been mutated, revealed that miR-141 regulates KEAP1 upon exposure to cisplatin. Finally, we show that the NF-κB pathway, which can be regulated by KEAP1, is activated upon miR-141 overexpression, and that inhibition of this pathway partially reverses miR-141-mediated cisplatin resistance. These findings demonstrate that the miR-141-mediated regulation of KEAP1 has a crucial role in the cellular response to cisplatin.

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Acknowledgements

We thank Dr Kevin Scanlon (Keck Graduate Institute of Applied Life Sciences, Claremont, USA) for the kind gift of the cisplatin-sensitive/resistant A2780 cell line pair. We thank Mariël Brok and Kirsten Ruigrok-Ritstier for technical assistance, Andrea Sacchetti for FACS-sorting KEAP1-transfected cells and Herman Burger for critical comments on the manuscript. This project is supported by a grant from the Dutch Cancer Society EMCR 2007-3794.

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Authors and Affiliations

  1. Department of Medical Oncology, Erasmus University Medical Center—Daniel den Hoed Cancer Center, Rotterdam, The Netherlands

    M T M van Jaarsveld, J Helleman, A W M Boersma, P F van Kuijk, R H J Mathijssen, E M J J Berns, J Verweij & E A C Wiemer

  2. Erasmus Center for Biomics, Erasmus MC, Rotterdam, The Netherlands

    W F van IJcken

  3. Department of Obstetrics and Gynecology and Leuven Cancer Institute, Division of Gynecologic Oncology, University Hospitals Leuven, Leuven, European Union, Belgium

    E Despierre & I Vergote

  4. Department of Genetics, Erasmus MC, Rotterdam, The Netherlands

    J Pothof

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van Jaarsveld, M., Helleman, J., Boersma, A. et al. miR-141 regulates KEAP1 and modulates cisplatin sensitivity in ovarian cancer cells. Oncogene 32, 4284–4293 (2013). https://doi.org/10.1038/onc.2012.433

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