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Phenazine derivatives attenuate the stemness of breast cancer cells through triggering ferroptosis

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Abstract

Breast cancer stem cells (BCSCs) are positively correlated with the metastasis, chemoresistance, and recurrence of breast cancer. However, there are still no drugs targeting BCSCs in clinical using for breast cancer treatment. Here, we tried to screen out small-molecule compounds targeting BCSCs from the phenazine library established by us before. We focused on the compounds without affecting cell viability and screened out three potential compounds (CPUL119, CPUL129, CPUL149) that can significantly attenuate the stemness of breast cancer cells, as evident by the decrease of stemness marker expression, CD44+/CD24– subpopulation, mammary spheroid-formation ability, and tumor-initiating capacity. Additionally, these compounds suppressed the metastatic ability of breast cancer cells in vitro and in vivo. Combined with the transcriptome sequencing analysis, ferroptosis was shown on the top of the most upregulated pathways by CPUL119, CPUL129, and CPUL149, respectively. Mechanistically, we found that these three compounds could trigger ferroptosis by accumulating and sequestering iron in lysosomes through interacting with iron, and by regulating the expression of proteins (IRP2, TfR1, ferritin) engaged in iron transport and storage. Furthermore, inhibition of ferroptosis rescued the suppression of these three compounds on breast cancer cell stemness. This study suggests that CPUL119, CPUL129, and CPUL149 can specifically inhibit the stemness of breast cancer cells through triggering ferroptosis and may be the potential compounds for breast cancer treatment.

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Abbreviations

BCSCs:

Breast cancer stem cells

CSCs:

Cancer stem cells

LSCs:

Leukemia stem cells

FDA:

Food and Drug Administration

ROS:

Reactive oxygen species

IRP2:

Iron-responsive element-binding protein 2

TfR1:

Transferrin receptor 1

IRE:

Iron regulatory element

FPN:

Ferroportin

GSEA:

Gene Set Enrichment Analysis

EMT:

Epithelial–mesenchymal transition

Taxol:

Paclitaxel

Adr:

Adriamycin

GSH:

Glutathione

FAC:

Ferric ammonium citrate

NMR:

Nuclear Magnetic Resonance

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Funding

This work was supported by the Project Program of National Nature Science Foundation of China (Grant No. 82173842, 81872757), Nature Science Foundation of Jiangsu Province of China (Grant No. BK20201329), the Fundamental Research Funds for the Central Universities (grant No. 2632018ZD01, No. 2632020ZD10), Innovation and Entrepreneurship Training Program for Undergraduate (No. 201910316220, No. 202010316245), the Medical Science and Technology Research Project of Henan Province (no. SBGJ202003010), the Science and Technology Research Project of Henan Province (no. 202102310158) and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

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Author notes

  1. Yue Yang, Yuanyuan Lu, and Chunhua Zhang contributed equally to this work.

Authors and Affiliations

  1. School of Life Science and Technology, School of Engineering, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 639 Longmian Road, Nanjing, 211198, People’s Republic of China

    Yue Yang, Yuanyuan Lu, Chunhua Zhang, Ting Wang, Zhuolu Xia, Jing Liu, Xiangyu Cheng, Tao Xi, Feng Jiang & Lufeng Zheng

  2. Department of Pharmacy, Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, 127 Dongming Road, Zhengzhou, 450003, People’s Republic of China

    Qianqian Guo & Wenzhou Zhang

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  1. Yue Yang
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  2. Yuanyuan Lu
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  3. Chunhua Zhang
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  10. Tao Xi
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  11. Feng Jiang
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Contributions

LZ, FJ and TX designed the research. YL and FJ approved the compounds. YY, CZ, ZX, JL, XC, TW, QG analyzed the data. YY, CZ performed the research. YY and LZwrote the paper. All authors read and approved the final manuscript.

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Correspondence to Tao Xi, Feng Jiang or Lufeng Zheng.

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All the experiments were obtained the approval of the Ethics Committee for Animal Experimentation of China Pharmaceutical University.

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Yang, Y., Lu, Y., Zhang, C. et al. Phenazine derivatives attenuate the stemness of breast cancer cells through triggering ferroptosis. Cell. Mol. Life Sci. 79, 360 (2022). https://doi.org/10.1007/s00018-022-04384-1

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