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SNAIL is induced by tamoxifen and leads to growth inhibition in invasive lobular breast carcinoma

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Abstract

Purpose

Invasive lobular carcinoma (ILC) is a histological subtype of breast cancer that is predominantly estrogen receptor alpha (ER)-positive (+) and is thus treated with endocrine therapies. Herein, we sought to understand the molecular underpinnings of the 4-hydroxytamoxifen (4OHT) resistance in ILC by assessing the potential role of the epithelial-to-mesenchymal transition transcription factor (EMT-TF) SNAIL (SNAI1).

Methods

Using a series of breast cancer cell lines, we measured the basal, estrogen and 4OHT-induced expression of SNAIL and other EMT-TF family members by quantitative reverse transcription-polymerase chain reaction and immunoblotting. Chromatin immunoprecipitation experiments were performed to assess ER binding to the SNAIL promoter. Cell proliferation, cell cycle and apoptosis were assessed in 2D cultures. 3D growth was assessed in Matrigel and Collagen I cultures.

Results

Estrogen and 4OHT induced SNAIL expression, but not that of the other EMT-TF family members SLUG (SNAI2) and SMUC (SNAI3), with the 4OHT effect being specific to the lobular but not the ductal subtype. We observed estrogen and 4OHT-induced ER recruitment to the SNAI1 promoter and high endogenous basal levels of SNAIL and several EMT-TFs in ILC cell lines. While SNAIL knockdown had a minor impact on the 4OHT partial agonism in estrogen-depleted conditions, it led to a surprising increase in cell proliferation in full serum. In complementary experiments, inducible SNAI1 overexpression caused decreased proliferation, associated with a cell cycle arrest in G0/G1. Additionally, apoptosis was observed in BCK4 cells.

Conclusion

These data suggest a previously unrecognized role for SNAIL in ILC, substantiating a context-dependent behavior for this EMT-TF.

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Acknowledgements

The authors thank Donald DeFranco, Ph.D., Adrian Lee, Ph.D., and Carola Neumann, M.D., at the University of Pittsburgh, as well as Adam Feinberg, Ph.D. at Carnegie Melon University for intellectual discussion and guidance with this work, and Jennifer Xavier, Ph.D. at the University of Pittsburgh for her critical reading and editing of the manuscript. Additionally, the authors extend gratitude to the laboratories of Adrian V. Lee, Carola Neumann, Mei Zhang, Yi Huang and Steffi Oesterreich in the Women’s Cancer Research Center at the University of Pittsburgh for their support and suggestions.

Funding

Research funding for this project was provided in part by a Susan G. Komen Scholar award (SO; SAC160073) and the Breast Cancer Research Foundation (SO). EAB was supported by a Department of Pharmacology and Chemical Biology John S. Lazo Fellowship, and a National Institutes of Health Ruth L. Kirschstein award [1F31CA203055-01]. NT was supported by a Department of Defense Breakthrough Fellowship Award [BC160764]. MJS was supported by a Department of Defense Postdoctoral Fellowship [BC110619], and a National Institutes of Health Pathway to Independence award [K99 CA193734]. KL was supported by a National Institutes of Health Ruth L. Kirschstein award [1F30CA203154-01]. TFB has received research funding for EMT studies from an American Lung Association Award (LCD 257864), V Foundation Scholar Award, Sidney Kimmel Foundation (SKF-15-099) and a Doris Duke Charitable Foundation Clinical Scientist Award (2015097), a Pittsburgh LUNG SPORE CDA P50CA090440 and American Cancer Society Research Scholar Grant (132939-RSG-18-185-01 TBG). This project used UPMC Hillman Cancer Center Core Facilities, supported by and NCI award [P30CA047904].

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Author notes

  1. Emily A. Bossart and Nilgun Tasdemir are equal contributors to this study.

Authors and Affiliations

  1. Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA

    Emily A. Bossart, Nilgun Tasdemir, Timothy F. Burns & Steffi Oesterreich

  2. Women’s Cancer Research Center, Magee-Womens Research Institute, University of Pittsburgh Cancer Institute, 204 Craft Avenue, Pittsburgh, PA, 15213, USA

    Emily A. Bossart, Nilgun Tasdemir, Amir Bahreini, Kevin M. Levine, Jian Chen, Ahmed Basudan & Steffi Oesterreich

  3. Department of Pathology, University of Colorado Anschutz Medical Campus, Denver, CO, 80045, USA

    Matthew J. Sikora & Britta M. Jacobsen

  4. Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Isfahan Province, Iran

    Amir Bahreini

  5. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA

    Kevin M. Levine

  6. Department of Clinical Laboratory Sciences, King Saud University, Riyadh, 11451, Saudi Arabia

    Ahmed Basudan

  7. Department of Medicine, UPMC Hillman Cancer Center, Pittsburgh, PA, 15232, USA

    Timothy F. Burns

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  1. Emily A. Bossart
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Correspondence to Steffi Oesterreich.

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Conflict of interest

Dr. Timothy F. Burns serves on an Advisory Board for AbbVie, Inc., and Dr. Oesterreich is a member of the External Scientific Advisory Board Panel of NSABP. The authors declare that they have no conflict of interest.

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Bossart, E.A., Tasdemir, N., Sikora, M.J. et al. SNAIL is induced by tamoxifen and leads to growth inhibition in invasive lobular breast carcinoma. Breast Cancer Res Treat 175, 327–337 (2019). https://doi.org/10.1007/s10549-019-05161-8

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