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Effect of bisphosphonates and statins on the in vitro radiosensitivity of breast cancer cell lines

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Abstract

Background

Early-stage breast cancer is usually treated with breast-conserving surgery followed by adjuvant radiation therapy. Acute skin toxicity is a common radiation-induced side effect experienced by many patients. Recently, a combination of bisphosphonates (zoledronic acid) and statins (pravastatin), or ZOPRA, was shown to radio-protect normal tissues by enhancing DNA double-strand breaks (DSB) repair mechanism. However, there are no studies assessing the effect of ZOPRA on cancerous cells. The purpose of this study is to characterize the in vitro effect of the zoledronic acid (ZO), pravastatin (PRA), and ZOPRA treatment on the molecular and cellular radiosensitivity of breast cancer cell lines.

Materials

Two breast cancer cell lines, MDA MB 231 and MCF-7, were tested. Cells were treated with different concentrations of pravastatin (PRA), zoledronate (ZO), as well as their ZOPRA combination, before irradiation. Anti-γH2AX and anti-pATM immunofluorescence were performed to study DNA DSB repair kinetics. MTT assay was performed to assess cell proliferation and viability, and flow cytometry was performed to analyze the effect of the drugs on the cell cycle distribution. The clonogenic assay was used to assess cell survival.

Results

ZO, PRA, and ZOPRA treatments were shown to increase the residual number of γH2AX foci for both cell lines. ZOPRA treatment was also shown to reduce the activity of the ATM kinase in MCF-7. ZOPRA induced a significant decrease in cell survival for both cell lines.

Conclusions

Our findings show that pretreatment with ZOPRA can decrease the radioresistance of breast cancer cells at the molecular and cellular levels. The fact that ZOPRA was previously shown to radioprotect normal tissues, makes it a good candidate to become a therapeutic window-widening drug.

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Data availability

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

Abbreviations

ATM:

Ataxia-telangiectasia mutated

pATM:

Phosphorylated ATM

DSB:

Double-strand breaks

DAPI:

4’,6’Diamidino-2-Phenyl-indole

Gy:

Gray (absorbed dose)

γH2AX:

Phosphorylated histone H2AX

IF:

Immunofluorescence

MN:

Micronuclei

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NHEJ:

Non-homologous end-joining

PBS:

Phosphate-buffered saline

PE:

Plating efficiency

PRA:

Pravastatin

RT:

Radiotherapy

SF:

Surviving fraction

ZO:

Zoledronic acid

ZOPRA:

Zoledronic acid and pravastatin

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Acknowledgements

We would like to thank all members of the Radiation Oncology department of AUBMC, Abou-Kheir’s Laboratory for their support and all members of the core facilities in the DTS Building for their help and support.

Funding

This research was supported by funding from the Medical Practice Plan of the American University of Beirut. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. Larry Bodgi and Jolie Bou-Gharios have been contributed equally.

  2. Wassim Abou-Kheir and Zeina Ayoub contributed equally as co-senior and co-corresponding authors.

Authors and Affiliations

  1. Department of Radiation Oncology, American University of Beirut Medical Center, Beirut, Lebanon

    Larry Bodgi, Jolie Bou-Gharios, Joyce Azzi, Rafka Challita, Charbel Feghaly, Fatima Chhade, Fady Geara & Zeina Ayoub

  2. Department of Anatomy, Cell Biology and Physiological Sciences, Faculty of Medicine, American University of Beirut, Beirut, Lebanon

    Larry Bodgi, Khanom Baalbaki, Hussein Kharroubi & Wassim Abou-Kheir

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  1. Larry Bodgi
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Contributions

Conceptualization, LB, WA, ZA; data acquisition and methodology, LB, JBG, JA, RC, CF, KB, HK, FC; validation and data analysis, LB, JBG, JA, FG, WA, ZA; writing and original draft preparation, LB, JBG; review and editing: all the authors. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Wassim Abou-Kheir or Zeina Ayoub.

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Bodgi, L., Bou-Gharios, J., Azzi, J. et al. Effect of bisphosphonates and statins on the in vitro radiosensitivity of breast cancer cell lines. Pharmacol. Rep 76, 171–184 (2024). https://doi.org/10.1007/s43440-023-00560-7

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