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.

LIU Xiao-dan;SONG Cheng-yang;KONG Cui-cui;TIAN Xin

Department of Breast Surgery,the Fourth Affiliated Hospital of China Medical University,Shenyang 110032,China;Department of Thoracic Surgery,the Fourth Affiliated Hospital of China Medical University,Shenyang 110032,China;Molecular Oncology Laboratory of Cancer Research Institute,the First Affiliated Hospital of China Medical University,Shenyang 110001,China

中国结合医学杂志（英文版）

[1]LIU Xiao-dan;SONG Cheng-yang;KONG Cui-cui;TIAN Xin-.Bufalin Induces Programmed Necroptosis in Triple-Negative Breast Cancer Drug-Resistant Cell Lines through RIP1/ROS-Mediated Pathway)[J].中国结合医学杂志（英文版）,2022(10):900-908

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