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Twist activates miR-22 to suppress estrogen receptor alpha in breast cancer

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Abstract

TWIST1 (Twist) is a basic helix-loop-helix transcription factor that is overexpressed in many cancers and promotes tumor cell invasion, metastasis, and recurrence. In this study, we demonstrate that Twist upregulates expression of microRNA 22 (miR-22) which, in turn, downregulates estrogen receptor alpha (ER) expression in breast cancer. Initial analysis of miR-22 and Twist expression in a panel of breast cancer cell lines showed a direct correlation between Twist and miR-22 levels with miR-22 being highly expressed in ER negative cell lines. Overexpressing Twist caused increased miR-22 levels while downregulating it led to decreased miR-22 expression. To characterize the upstream promoter region of miR-22, we utilized rapid amplification of cDNA ends and identified the transcription start site and the putative promoter region of miR-22. Mechanistically, we determined that Twist, in combination with HDAC1 and DNMT3B, transcriptionally upregulates miR-22 expression by binding to E-boxes in the proximal miR-22 promoter. We also established that miR-22 causes an increase in growth in 3D but not 2D cultures. Importantly, we observed a direct correlation between increased breast cancer grade and Twist and miR-22 expression. We also identified two potential miR-22 binding sites in the 3′-UTR region of ER and confirmed by promoter assays that miR-22 regulates ER expression by binding to both target sites. These results reveal a novel pathway of ER suppression by Twist through miR-22 activation that could potentially promote the ER negative phenotype in breast cancers.

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Acknowledgements

We would like to acknowledge John Domek, Ashley Irving, and Yehudit Bergman for technical help with this work.

Funding

This work was supported by the Maryland Stem Cell Research Fund [MDTSCRC0072 to F.V., MDTSCRC0064 to V.R.] and the National Institutes of Health [R01CA097226 to V.R.].

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Authors and Affiliations

  1. Department of Radiology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    Farhad Vesuna, Ala Lisok & Venu Raman

  2. Department of Pathology, University of Utrecht, Utrecht, The Netherlands

    Paul van Diest

  3. Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA

    Venu Raman

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  1. Farhad Vesuna
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  2. Ala Lisok
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  3. Paul van Diest
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  4. Venu Raman
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Contributions

FV performed the investigation, conceptualization, methodology, analysis, and writing - original draft preparation. AL performed investigation. PvD contributed to the supply of breast cancer samples. VR performed supervision, reviewing, and editing.

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Correspondence to Farhad Vesuna or Venu Raman.

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We used anonymous archival leftover pathology material sourced from Utrecht Medical School. No ethical approval is required as use of de-identified leftover material is part of the standard agreement with patients in the hospital.

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Vesuna, F., Lisok, A., van Diest, P. et al. Twist activates miR-22 to suppress estrogen receptor alpha in breast cancer. Mol Cell Biochem 476, 2295–2306 (2021). https://doi.org/10.1007/s11010-021-04065-w

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