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The effect of hesperetin on estrogen receptor gene expression and its relationship with the downstream pathways of estrogen receptor alpha

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Abstract

Background

Estrogen receptor (ER) is a transcription factor that affects the expression of some genes involved in the progression and development of breast cancer (BC). Hesperetin (Hst) is a flavonoid that inhibits the proliferation of BC cells. In this study, we investigated the effect of Hst on the cell viability of MCF-7 cells and the gene expression of the ERα, ERβ, IL-6, Ps2, and Cyclin D1.

Methods

In this study, cell viability was determined by MTT assay. The cells were seeded in RPMI-1640 medium and then exposed to different concentrations of Hst (0, 25, 50, 100, 200, and 400 µM) for 24 h, and IC50 was calculated. Real-time PCR was used to assess the expression of ERα, ERβ, pS2, Cyclin D1, and IL-6 mRNA. MCF-7 cells were seeded in RPMI-1640 medium and then exposed to different concentrations of Hst (0, 25, 50, 100, and 200 µM) for 24 h. Real-time PCR was carried out using a Step One Real-Time PCR System (ABI, USA) and Amplicon SYBR Green reagents.

Results

The MTT assay revealed increased cytotoxicity with higher concentrations of Hst, and the IC50 was calculated at 200 µM. Real-time PCR analysis following treatment with Hst showed a significant increase in ERα gene expression at 25 µM of Hst and a decrease in expression at 50, 100, and 200 µM of Hst (p < 0.0001). ERβ gene expression significantly decreased across all concentrations of Hst (p < 0.0001), while IL-6 gene expression decreased significantly in all concentrations (p < 0.0001). pS2 gene expression increased significantly with all concentrations of Hst (p < 0.0001), while Cyclin D1 gene expression did not significantly decrease upon Hst exposure (p > 0.05).

Conclusions

The results of our study demonstrate that Hst has the ability to induce cell death in MCF-7 cells. Furthermore, it was observed that Hst reduces the expression of the ER gene and enhances its activity, which can affect the downstream pathways of the ER.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by Birjand University of medical sciences.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran

    Milad Bideh, Samaneh Safari & Mohammad Zangooei

  2. Department of Biochemistry, Faculty of Medicine, Ahvaz Jundishapur University of Medical sciences, Ahvaz, Iran

    Azam Khedri

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  1. Milad Bideh
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  2. Samaneh Safari
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  4. Mohammad Zangooei
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Contributions

MZ contributed to the study’s conception and design. MB performed material preparation, data collection, and analysis. SS wrote the first draft of the manuscript and all authors commented on previous versions. MZ and AZ read and approved the final manuscript.

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Correspondence to Mohammad Zangooei.

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The authors have no relevant financial or non-financial interests to disclose.

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This is cell line-based study. No ethical approval is required.

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Bideh, M., Safari, S., Khedri, A. et al. The effect of hesperetin on estrogen receptor gene expression and its relationship with the downstream pathways of estrogen receptor alpha. Mol Biol Rep 50, 7225–7236 (2023). https://doi.org/10.1007/s11033-023-08616-w

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