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Zoledronic acid induces ferroptosis by upregulating POR in osteosarcoma

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Abstract

Osteosarcoma, usually originating in the stroma, is the most common primary bone cancer in adolescents, and its prognosis is poor. Surgery, adjuvant and neoadjuvant chemotherapy and radiation therapy are not satisfactory at the present time. Therefore, it is critical to develop novel therapeutic strategies to improve the quality of life and long-term survival rate of osteosarcoma patients. In this study, we discovered that zoledronic acid (ZOL) dramatically increased cell death in osteosarcoma cells, and this cytotoxicity was greatly reversed by liproxstatin-1 (a ferroptosis inhibitor). ZOL also had an obvious effect on lipid peroxidation and reactive oxygen species (ROS), which suggested that ZOL most certainly induces ferroptosis in osteosarcoma cells. In addition, we further found that ZOL increases cytochrome P450 oxidoreductase (POR) expression dose dependently in osteosarcoma cell lines. Knockdown of POR attenuated ZOL-induced cytotoxicity and attenuated the effect of ferroptosis in osteosarcoma cells, which indicated that POR plays an important role in ferroptosis. Moreover, we also found that ZOL inhibits osteosarcoma growth in vivo. Our findings suggest that ZOL induces ferroptosis by upregulating POR expression to increase ROS levels and upregulate lipid peroxidation levels in osteosarcoma cells. POR may be used as a therapeutic target to inhibit osteosarcoma.

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Data availability

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

Abbreviations

ZOL:

Zoledronic acid

ROS:

Reactive oxygen species

DEGs:

Differentially expressed genes

KEGG:

Kyoto Encyclopedia of Genes and Genomes

GO:

Gene Ontology

RIPA:

Radioimmunoprecipitation assay

FITC:

Fluorescein isothiocyanate

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Acknowledgements

Not applicable.

Funding

This study was supported by the National Natural Science Foundation of China (81760041) and the Shenzhen Science and Technology Program (JCYJ20220530144201002).

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Authors and Affiliations

  1. Department of Orthopedics, The Eighth Affiliated Hospital, Sun Yat-Sen University, No. 3025, Shennan Middle Road, Futian District, Shenzhen, 518033, Guangdong, People’s Republic of China

    Hong Jiacong, Li Haonan, Song Yichang & Chen Keng

  2. Department of Hematology, The Eighth Affiliated Hospital, Sun Yat-Sen University, No. 3025, Shennan Middle Road, Futian District, Shenzhen, 518033, Guangdong, People’s Republic of China

    Yang Qirui & Chen Yan

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  1. Hong Jiacong
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Contributions

HJC: the conception and design of the study, drafting the article; YQR: analysis of data; LHN and Song Yichang: interpretation and acquisition of data; CK and CY: revising article, final approval of the version to be submitted. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Chen Yan or Chen Keng.

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Conflict of interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Ethical approval

The study was approved by the Ethics Committee of The Eighth Affiliated Hospital of Sun Yat-sen University. Animal Studies was supported by The Eighth Affiliated Hospital of Sun Yat-sen University. All animal experiments were conducted in accordance with the National Institutes of Health Laboratory Animal Care and Use Guidelines.

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12032_2023_1988_MOESM1_ESM.tif

Supplementary file1 (TIF 27405 KB)—A, Colony formation capacity of MG63 and 143B cells, as assessed by a colony formation assay after 10 days. The colony formation assay showed ZOL inhibit the osteosarcoma cells. B, ZOL dose-dependently decreased the cell viability of osteosarcoma. C, Cell apoptosis were detected by flow cytometry which showed that ZOL can’ t cause apoptosis in OS cells. D, Western blot analysis showed that groups' protein expression levels appeared to be similar, which showed that ZOL can’ t induce autophagy in OS cells. E, After 2 weeks, the ZOL group and NC group which were subcutaneously injected with ZOL-treated 143B cells or normal 143B cells respectively both successfully formed xenograft tumors. (Every circle means each nude mice. Yellow means xenograft tumor has been formed, grey means xenograft tumor has not been formed) F, The change of tumor volume showed ZOL could inhibit the growth and proliferation of osteosarcoma. *P <0.05; **P <0.01

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Jiacong, H., Qirui, Y., Haonan, L. et al. Zoledronic acid induces ferroptosis by upregulating POR in osteosarcoma. Med Oncol 40, 141 (2023). https://doi.org/10.1007/s12032-023-01988-w

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