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RANK-c attenuates aggressive properties of ER-negative breast cancer by inhibiting NF-κB activation and EGFR signaling

Oncogene volume 37, pages 5101–5114 (2018)Cite this article

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Abstract

The RANK/RANKL axis emerges as a key regulator of breast cancer initiation, progression, and metastasis. RANK-c is a RANK receptor isoform produced through alternative splicing of the TNFRSF11A (RANK) gene and a dominant-negative regulator of RANK-induced nuclear factor-κB (NF-κB) activation. Here we report that RANK-c transcript is expressed in 3.2% of cases in The Cancer Genome Atlas breast cancer cohort evenly between ER-positive and ER-negative cases. Nevertheless, the ratio of RANK to RANK-c (RANK/RANK-c) is increased in ER-negative breast cancer cell lines compared to ER-positive breast cancer cell lines. In addition, forced expression of RANK-c in ER-negative breast cancer cell lines inhibited stimuli-induced NF-κB activation and attenuated migration, invasion, colony formation, and adhesion of cancer cells. Further, RANK-c expression in MDA-MB-231 cells inhibited lung metastasis and colonization in vivo. The RANK-c-mediated inhibition of cancer cell aggressiveness and nuclear factor-κB (NF-κB) activation in breast cancer cells seems to rely on a RANK-c/TNF receptor-associated factor-2 (TRAF2) protein interaction. This was further confirmed by a mutated RANK-c that is unable to interact with TRAF2 and abolishes the ability to attenuate NF-κB activation, migration, and invasion. Additional protein interaction characterization revealed epidermal growth factor receptor (EGFR) as a novel interacting partner for RANK-c in breast cancer cells with a negative effect on EGFR phosphorylation and EGF-dependent downstream signaling pathway activation. Our findings further elucidate the complex molecular biology of the RANKL/RANK system in breast cancer and provide preliminary data for RANK-c as a possible marker for disease progression and aggressiveness.

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Acknowledgements

We thank the Advanced Light Microscopy facility of the Medical School, University of Patras, Greece. We also thank Dr. Ross (Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, MA 01605, USA) for plasmid MSCV-XZ066-EGFRvIII (Addgene #20737).

Funding

CS is funded by the State Scholarships Foundation “IKY Fellowships of Excellence for Postgraduate Studies in Greece-Siemens Programme” (SR 229101). The research reported was partially funded by the State Scholarships Foundation (SR 229101). The publicly available dataset from the TCGA breast cancer study was retrieved from the TCGA data portal on 05/2017.

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Author notes

  1. These authors contributed equally: Chaido Sirinian, Anastasios D. Papanastasiou.

Authors and Affiliations

  1. Clinical and Molecular Oncology Laboratory, Division of Oncology, Department of Medicine, University of Patras, Patras, Greece

    Chaido Sirinian, Anastasios D. Papanastasiou & Haralabos P. Kalofonos

  2. Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    Michail Schizas & Tari A. King

  3. Laboratory for Molecular Respiratory Carcinogenesis, Department of Physiology, Faculty of Medicine, University of Patras, Patras, Greece

    Magda Spella & Georgios T. Stathopoulos

  4. Department of Pathology, Patras General Hospital, Patras, Greece

    Maria Repanti

  5. Department of Biology, School of Medicine, University of Patras, Patras, Greece

    Ioannis K. Zarkadis

  6. Surgical Oncology, Brigham and Women’s Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA

    Tari A. King

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Sirinian, C., Papanastasiou, A.D., Schizas, M. et al. RANK-c attenuates aggressive properties of ER-negative breast cancer by inhibiting NF-κB activation and EGFR signaling. Oncogene 37, 5101–5114 (2018). https://doi.org/10.1038/s41388-018-0324-y

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