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ZEB2 regulates endocrine therapy sensitivity and metastasis in luminal a breast cancer cells through a non-canonical mechanism

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Abstract

Purpose

The transcription factors ZEB1 and ZEB2 mediate epithelial-to-mesenchymal transition (EMT) and metastatic progression in numerous malignancies including breast cancer. ZEB1 and ZEB2 drive EMT through transcriptional repression of cell–cell junction proteins and members of the tumor suppressive miR200 family. However, in estrogen receptor positive (ER +) breast cancer, the role of ZEB2 as an independent driver of metastasis has not been fully investigated.

Methods

In the current study, we induced exogenous expression of ZEB2 in ER + MCF-7 and ZR-75–1 breast cancer cell lines and examined EMT gene expression and metastasis using dose–response qRT-PCR, transwell migration assays, proliferation assays with immunofluorescence of Ki-67 staining. We used RNA sequencing to identify pathways and genes affected by ZEB2 overexpression. Finally, we treated ZEB2-overexpressing cells with 17β-estradiol (E2) or ICI 182,780 to evaluate how ZEB2 affects estrogen response.

Results

Contrary to expectation, we found that ZEB2 did not increase canonical epithelial nor decrease mesenchymal gene expressions. Furthermore, ZEB2 overexpression did not promote a mesenchymal cell morphology. However, ZEB1 and ZEB2 protein expression induced significant migration of MCF-7 and ZR-75-1 breast cancer cells in vitro and MCF-7 xenograft metastasis in vivo. Transcriptomic (RNA sequencing) pathway analysis revealed alterations in estrogen signaling regulators and pathways, suggesting a role for ZEB2 in endocrine sensitivity in luminal A breast cancer. Expression of ZEB2 was negatively correlated with estrogen receptor complex genes in luminal A patient tumors. Furthermore, treatment with 17β-estradiol (E2) or the estrogen receptor antagonist ICI 182,780 had no effect on growth of ZEB2-overexpressing cells.

Conclusion

ZEB2 is a multi-functional regulator of drug sensitivity, cell migration, and metastasis in ER + breast cancer and functions through non-canonical mechanisms.

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Acknowledgements

Thank you to Dr. Van T. Hoang, PhD and the animal vivarium staff at Tulane University School of Medicine for their assistance with the in vivo experiments. High throughput sequencing was performed at the Center for Genomics and Bioinformatics at Indiana University, Bloomington.

Funding

This project was funded by the National Institutes of Health R01-CA174785-01A1 (BMC-B) and the National Institutes of Health R01-CA125806-02 (MEB).

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Author notes

  1. Hope E. Burks and Margarite D. Matossian have contributed equally and should be considered co-first authors.

Authors and Affiliations

  1. Department of Medicine, Section of Hematology & Medical Oncology, Tulane University Health Sciences Center, New Orleans, LA, 70112, USA

    Hope E. Burks, Margarite D. Matossian, Steven Elliott, Bridgette M. Collins-Burow & Matthew E. Burow

  2. Tulane Cancer Center, New Orleans, LA, 70112, USA

    Hope E. Burks, Margarite D. Matossian, Steven Elliott, Bridgette M. Collins-Burow & Matthew E. Burow

  3. Florida Gulf Coast University, Fort Myers, FL, 33965, USA

    Lyndsay Vanhoy Rhodes

  4. Department of Physics, Tulane University, New Orleans, LA, 70112, USA

    Theresa Phamduy & Douglas Chrisey

  5. Center of Genomics and Bioinformatics, Indiana University, Bloomington, IN, 47405, USA

    Aaron Buechlein, Douglas B. Rusch & David F. B. Miller

  6. Medical Sciences Program, Indiana University School of Medicine, Bloomington, IN, 47405, USA

    David F. B. Miller & Kenneth P. Nephew

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  1. Hope E. Burks
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Corresponding author

Correspondence to Matthew E. Burow.

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The authors have no conflicts of interest to disclose.

Ethical approval

All procedures involving these animals were conducted in compliance with State and Federal laws, standards of the U.S. Department of Health and Human Services, and guidelines established by the Tulane University Animal Care and Use Committee. The Tulane University Animal Care and Use Committee approved the use of animals in this specific study. The facilities and laboratory animal program of Tulane University are accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care.

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Burks, H.E., Matossian, M.D., Rhodes, L.V. et al. ZEB2 regulates endocrine therapy sensitivity and metastasis in luminal a breast cancer cells through a non-canonical mechanism. Breast Cancer Res Treat 189, 25–37 (2021). https://doi.org/10.1007/s10549-021-06256-x

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