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Introduction of hsa-miR-512-3p as a new regulator of HER2 signaling pathway in breast cancer

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Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Purpose

Dysregulation of HER2 signaling pathway in breast cancer is well documented. Our bioinformatics analysis predicted hsa-miR-512-3p (miR-512-3p) as a bona fide regulator of HER2 as well as HER3, PIK3R2, and AKT1 genes. Then, we intended to examine the effect of miR-512-3p on the predicted target genes that are involved in HER2 signaling pathway.

Methods and results

RT-qPCR results indicated lower expression of miR-512-3p in breast cancer specimens, compared to their normal pairs. Overexpression of miR-512-3p resulted in HER2, HER3, PIK3R2, and AKT1 gene downregulation, detected by RT-qPCR and the result was confirmed by western analysis and ELIZA test against p-AKT, BAX, FADD, and HER2 proteins in SKBR3 cells, respectively. Then, dual-luciferase assay supported the direct interaction of miR-512-3p with 3′UTR sequences of HER2, HER3, PIK3R2, and AKT1 target genes. When miR-512-3p was overexpressed, BAX/BCL2 expression ratio and proportion of sub-G1 cell population were increased in transfected SKBR3 cells, detected by RT-qPCR and flow cytometry, respectively. These results were consistent with the decreased viability of transfected cells, documented by MTT assay. In addition, results were consistent with the upregulation of BAX, BAK, BOK, PTEN, P53, and P21 genes and downregulation of CCND1 gene in SKBR3 cells. Although the overexpression of miR-512 resulted in cell cycle arrest at Sub-G1 stage in MDA-MB-231 cells, this effect seemed independent of targeting HER2, HER3, PIK3R2, and AKT1 target genes.

Conclusion

Overall, results indicated that miR-512-3p acts as a cell-type-specific tumor suppressor, through targeting HER2, HER3, PIK3R2, and AKT1 transcripts. These results suggest miR-512-3p as a potential candidate marker for breast cancer diagnosis.

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Acknowledgements

We wish to thank 4402 Lab members at Tarbiat Modares University and Motamed Cancer Institute (MCI) for their supports throughout the project.

Funding

This study was funded by INSF (Grant Number 96012144).

Author information

Authors and Affiliations

  1. Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 111-14115, Iran

    Zahra Mohamadzade & Bahram M. Soltani

  2. Clinical Genetic Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

    Frouzande Mahjoubi

Authors
  1. Zahra Mohamadzade
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  2. Frouzande Mahjoubi
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  3. Bahram M. Soltani
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Contributions

ZM and BMS conceived and designed the study, analyzed the results, and wrote the paper. ZM performed the experiments. FM provided cancer tissue samples with preparation of pathological features. All authors reviewed the results and approved the final version of the manuscript.

Corresponding author

Correspondence to Bahram M. Soltani.

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Conflict of interest

The authors have no conflict of interest to declare.

Ethical approval

This study was approved by the Institutional ethics committee at TMU. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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Written informed consent was obtained from patients involved in the study.

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Mohamadzade, Z., Mahjoubi, F. & Soltani, B.M. Introduction of hsa-miR-512-3p as a new regulator of HER2 signaling pathway in breast cancer. Breast Cancer Res Treat 185, 95–106 (2021). https://doi.org/10.1007/s10549-020-05937-3

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  • DOI: https://doi.org/10.1007/s10549-020-05937-3

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