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CRISPR/Cas9 mediated knocking out of OPN gene enhances radiosensitivity in MDA-MB-231 breast cancer cell line

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Abstract

Purpose

Although chemotherapy and radiotherapy in conjunction with surgery have been known as the standard methods for patients with breast cancer, they frequently face resistance due to the failure of cells to death. Accordingly, improving the results requires discovering novel therapeutic approaches based on the changes in the molecular biology of cancer cells. Osteopontin (OPN) is a secreted protein that previous studies have shown to be associated with progression, poor prognosis, and metastasis in breast cancer. The current study examined the synergistic effects of radiotherapy and knocking out of OPN gene, utilizing CRISPR/Cas9 technique in MDA-MB-231 breast cancer cells.

Methods

We used to knock out the OPN gene by the two different gRNAs. The cells irradiated 24 h after transfection. The mRNA expression, tumor cell proliferation, cell cycle distribution, growth, and apoptosis were measured. Moreover, activation of Chk1 and AKT were measured via western blot.

Results

We demonstrated the OPN knocking out along with radiation led to the promotion of apoptosis, suppression of downstream genes, reduction of cell viability, and inhibition of cell-cycle progression. The western blot analysis has indicated that the knocking out of the OPN gene along with radiotherapy changes DNA damage responses substantially.

Conclusions

The OPN gene knocking out with radiotherapy might be an efficient approach to overcome the radioresistance in breast cancer.

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Data availability

The datasets analyzed during the current study are available from the first author [Rahil Ghanbarnasab Behbahani] on reasonable request.

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Funding

This study was supported by the Ahvaz Jundishapur University of Medical Sciences (grant No. U-96164). The research was planned for the M.Sc. thesis of Rahil Ghanbarnasab Behbahani.

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Authors and Affiliations

  1. Department of Medical Physics, School of Medicine, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran

    Rahil Ghanbarnasab Behbahani, Amir Danyaei & Mohammad Javad Tahmasbi

  2. Department of Virology, Faculty of Medicine, Hamedan University of Medical Sciences, Hamedan, Iran

    Ali Teimoori

  3. Department of Virology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Niloofar Neisi

Authors
  1. Rahil Ghanbarnasab Behbahani
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  2. Amir Danyaei
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  3. Ali Teimoori
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  4. Mohammad Javad Tahmasbi
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  5. Niloofar Neisi
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Contributions

All authors contributed to the study’s conception and design. Material preparation, data collection, and analysis were performed by RGHB, AD, AT, MJT, and NN. The first draft of the manuscript was written by RGHB and AD. All authors commented on previous versions of the manuscript and all authors read and approved the final manuscript.

Corresponding author

Correspondence to Amir Danyaei.

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Competing interests

The authors declare no competing interests.

Conflict of interest

The authors declare no competing interests.

Ethical approval

AJUMS’s Human Ethics Committee approved all protocols (IR.AJUMS.REC.1396.1009).

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Informed consent was obtained from all individual participants included in the study.

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Ghanbarnasab Behbahani, R., Danyaei, A., Teimoori, A. et al. CRISPR/Cas9 mediated knocking out of OPN gene enhances radiosensitivity in MDA-MB-231 breast cancer cell line. J Cancer Res Clin Oncol 149, 4117–4130 (2023). https://doi.org/10.1007/s00432-022-04304-7

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  • DOI: https://doi.org/10.1007/s00432-022-04304-7

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