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Role of circ-FOXO3 and miR-23a in radiosensitivity of breast cancer

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Abstract

Identifying the radiosensitivity of cells before radiotherapy (RT) in breast cancer (BC) patients allows appropriate switching between routinely used treatment regimens and reduces adverse side effects in exposed patients. In this study, blood was collected from 60 women diagnosed with Invasive Ductal Carcinoma (IDC) BC and 20 healthy women. To predict cellular radiosensitivity, a standard G2-chromosomal assay was performed. From these 60 samples, 20 BC patients were found to be radiosensitive based on the G2 assay. Therefore, molecular studies were finally performed on two equal groups (20 samples each) of patients with and without cellular radiosensitivity. QPCR was performed to examine the expression levels of circ-FOXO3 and miR-23a in peripheral blood mononuclear cells (PBMCs) and RNA sensitivity and specificity were determined by plotting Receiver Operating Characteristic (ROC) curves. Binary logistic regression was performed to identify RNA involvement in BC and cellular radiosensitivity (CR) in BC patients. Meanwhile, qPCR was used to compare differential RNA expression in the radiosensitive MCF-7 and radioresistant MDA-MB-231 cell lines. An annexin -V FITC/PI binding assay was used to measure cell apoptosis 24 and 48 h after 2 Gy, 4 Gy, and 8 Gy gamma-irradiation. Results indicated that circ-FOXO3 was downregulated and miR-23a was upregulated in BC patients. RNA expression levels were directly associated with CR. Cell line results showed that circ-FOXO3 overexpression induced apoptosis in the MCF-7 cell line and miR-23a overexpression inhibited apoptosis in the MDA-MB-231 cell line. Evaluation of the ROC curves revealed that both RNAs had acceptable specificity and sensitivity in predicting CR in BC patients. Binary logistic regression showed that both RNAs were also successful in predicting breast cancer. Although only circ-FOXO3 has been shown to predict CR in BC patients, circ-FOXO3 may function as a tumor suppressor and miR-23a may function as oncomiR in BC. Circ-FOXO3 and miR-23a may be promising potential biomarkers for BC prediction. Furthermore, Circ-FOXO3 could be a potential biomarker for predicting CR in BC patients.

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Acknowledgements

This research was supported by the research department of the Faculty of Medical Sciences of Tarbiat Modares University, Tehran, Iran. The authors sincerely appreciate the head and staff of the Oncology Department of Imam Khomeini Hospital for their valuable cooperation. The authors also thank all patients and healthy volunteers who participated in this study. We also thank Mr. H. Nosrati for the irradiation of blood and cell line samples.

Funding

This research was supported by a grant (Grant Number: IG-39711) from the research department of the Tarbiat Modares University, Tehran, Iran.

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Authors and Affiliations

  1. Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Elahe Abdollahi & Hossein Mozdarani

  2. Unit of Personalized Medicine, Department of Epidemiology, University Medical Center Groningen, Groningen, The Netherlands

    Behrooz Z. Alizadeh

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  1. Elahe Abdollahi
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  2. Hossein Mozdarani
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All authors contributed to the study's conception and design. Supervision and study design [HM]. Material preparation, data collection, and analysis were done by [EA], and [HM]. The first draft of the manuscript was written by [EA] and editing was performed by [BA and HM]. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Hossein Mozdarani.

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Abdollahi, E., Mozdarani, H. & Alizadeh, B.Z. Role of circ-FOXO3 and miR-23a in radiosensitivity of breast cancer. Breast Cancer 30, 714–726 (2023). https://doi.org/10.1007/s12282-023-01463-4

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