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Bufalin Induces Programmed Necroptosis in Triple-Negative Breast Cancer Drug-Resistant Cell Lines through RIP1/ROS-Mediated Pathway

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Abstract

Objective

To explore the effect and mechanism of action of bufalin in triple-negative breast cancer (TNBC) drug-resistant cell lines.

Methods

The normal human mammary epithelial cell line, TNBC cell line, TNBC adriamycin-resistant cell line, and TNBC docetaxel-resistant cell line were treated with different doses of bufalin (0–1,000 nmol/L) at different time points (0–72 h). Propidium iodide staining, AV-FITC/PI double staining, Hoechst 33342/PI double staining and transmission electron microscopy (TEM) were used to evaluate the death patterns of the cell lines.

Results

Bufalin killed the TNBC cell line and its drug-resistant cell lines in a dose/time-dependent manner (all P<0.01). After treatment with bufalin for 24 h, the adriamycin-resistant cell line showed a co-existing pattern of necroptosis and apoptosis. However, at 48 h, necroptosis was the main manifestation. After treatment with bufalin, the expressions of tumor necrosis factor α, phospho-tumor necrosis factor receptor 1, phospho-receptor interacting protein 1 and c-caspase 3 increased (all P<0.01), the killing effect of bufalin could be mostly inhibited by NEC-1, and by z-VAD-fmk (both P<0.01). Besides, the intracellular reactive oxygen species (ROS) levels increased considerably (P<0.01), the antioxidant N-acetyl cysteine or Nec-1 could inhibit the increase of ROS level and the killing effect of bufalin (all P<0.01). The adriamycin-resistant cell line exhibited necroptosis characteristic after 48 h of bufalin treatment under TEM.

Conclusions

Bufalin could induce necroptosis through RIP1/ROS-mediated pathway to kill the drug-resistant TNBC cell lines. This finding provides critical experimental data and theoretical basis for the clinical application of bufalin to overcome the difficulties in the treatment of TNBC.

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Author information

Authors and Affiliations

  1. Department of Breast Surgery, the Fourth Affiliated Hospital of China Medical University, Shenyang, 110032, China

    Xiao-dan Liu

  2. Department of Thoracic Surgery, the Fourth Affiliated Hospital of China Medical University, Shenyang, 110032, China

    Cheng-yang Song

  3. Molecular Oncology Laboratory of Cancer Research Institute, the First Affiliated Hospital of China Medical University, Shenyang, 110001, China

    Cui-cui Kong & Xin Tian

Authors
  1. Xiao-dan Liu
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  2. Cheng-yang Song
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  3. Cui-cui Kong
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  4. Xin Tian
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Contributions

Liu XD, Song CY and Kong CC performed the experiments; Tian X designed the experiments and drafted the manuscript. All authors read and approved the final version of the manuscript for publication.

Corresponding author

Correspondence to Xin Tian.

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Conflict of Interest

The authors declare no financial or commercial conflict of interest.

Supported by the National Natural Science Foundation of China (No. 81573654)

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Liu, Xd., Song, Cy., Kong, Cc. et al. Bufalin Induces Programmed Necroptosis in Triple-Negative Breast Cancer Drug-Resistant Cell Lines through RIP1/ROS-Mediated Pathway. Chin. J. Integr. Med. 28, 900–908 (2022). https://doi.org/10.1007/s11655-021-3458-7

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  • DOI: https://doi.org/10.1007/s11655-021-3458-7

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