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A novel interaction between HER2/neu and cyclin E in breast cancer

Oncogene volume 29, pages 3896–3907 (2010)Cite this article

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Abstract

HER2/neu (HER2) and cyclin E are important prognostic indicators in breast cancer. As both are involved in cell cycle regulation we analyzed whether there was a direct interaction between the two. HER2 and cyclin E expression levels were determined in 395 breast cancer patients. Patients with HER2-overexpression and high levels of cyclin E had decreased 5-year disease-specific survival compared with low levels of cyclin E (14% versus 89%, P<0.0001). In vitro studies were performed in which HER2-mediated activity in HER2-overexpressing breast cancer cell lines was downregulated by transfection with HER2 small interfering RNA or treatment with trastuzumab. Cyclin E expression levels were determined by western blot analysis, and functional effects analyzed using kinase assays, MTT assays were used to assess cell viability as a marker of proliferation and fluorescence-activated cell sorting analysis was used to determine cell cycle profiles. Decreased HER2-mediated signaling resulted in decreased expression of cyclin E, particularly the low molecular weight (LMW) isoforms. Decreased HER2 and LMW cyclin E expression had functional consequences, including decreased cyclin E-associated kinase activity and decreased proliferation, because of increased apoptosis and an increased accumulation of cells in the G1 phase. In vivo studies performed in a HER2-overexpressing breast cancer xenograft model confirmed the effects of trastuzumab on cyclin E expression. Given the relationship between HER2 and cyclin E, in vitro clonogenic assays were performed to assess combination therapy targeting both proteins. Isobologram analysis showed a synergistic interaction between the two agents (trastuzumab targeting HER2 and roscovitine targeting cyclin E). Taken together, these studies show that HER2-mediated signaling effects LMW cyclin E expression, which in turn deregulates the cell cycle. LMW cyclin E has prognostic and predictive roles in HER2-overexpressing breast cancer, warranting further study of its potential as a therapeutic target.

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Acknowledgements

This work was supported by NIH T32 CA009599 (Mittendorf); AACR (Mittendorf); Susan G Komen for the Cure PDF0707621 (Keyomarsi and Mittendorf); Clayton Foundation for Research (Keyomarsi); NIH CA87458 (Keyomarsi); NCI P50CA116199 (Keyomarsi); Susan G Komen Breast Cancer Foundation BCTR0504200 (Hunt) and NIH 1K99CA133244-01 (Mittendorf).

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Author notes

  1. K Keyomarsi and K K Hunt: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Surgical Oncology, MD Anderson Cancer Center, Houston, TX, USA

    E A Mittendorf, Y Liu, T McKenzie, N Qiao & K K Hunt

  2. Departments of Bioinformatics and Computational Biology, MD Anderson Cancer Center, Houston, TX, USA

    S L Tucker

  3. Department of Experimental Radiation Oncology, MD Anderson Cancer Center, Houston, TX, USA

    S Akli, A Biernacka, Y Liu, K Keyomarsi & K K Hunt

  4. Protein Phosphorylation and Disease, Centre National de la Recherche Scientifique, Station Biologique, Roscoff, France

    L Meijer

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  1. E A Mittendorf
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  2. Y Liu
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  3. S L Tucker
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Correspondence to K Keyomarsi or K K Hunt.

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Competing interests

Dr L Meijer is a coinventor on the patent on roscovitine licensed to Cyclacel.

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Mittendorf, E., Liu, Y., Tucker, S. et al. A novel interaction between HER2/neu and cyclin E in breast cancer. Oncogene 29, 3896–3907 (2010). https://doi.org/10.1038/onc.2010.151

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