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中国癌症防治杂志

中国癌症防治杂志 ›› 2023, Vol. 15 ›› Issue (3): 278-284.doi: 10.3969/j.issn.1674-5671.2023.03.05

• 基础研究 • 上一篇    下一篇

小檗碱诱导前列腺癌细胞铁死亡及其作用机制

  

  1. 南阳市第二人民医院泌尿男科
  • 出版日期:2023-06-25 发布日期:2023-06-19
  • 通讯作者: 郭新武 E?mail：564622681@qq.com
  • 基金资助:
    南阳市科技发展计划项目（KJGG2018124）

Berberine induces ferroptosis in prostate cancer cells and its mechanisms

  • Online:2023-06-25 Published:2023-06-19

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摘要： 目的 探讨小檗碱对前列腺癌细胞铁死亡的影响及其作用机制。方法 体外培养前列腺癌DU145和PC⁃3细胞，采用CCK⁃8检测不同浓度（1.560、3.125、6.250、12.500、25.000、50.000、100.000 μmol/L）小檗碱对前列腺癌细胞增殖活力的影响，并计算半数抑制浓度（half⁃inhibitory concentration，IC50）；采用谷胱甘肽（glutathione，GSH）试剂盒检测细胞内GSH水平；胱氨酸定量分析试剂盒检测细胞内胱氨酸水平，DCFH⁃DA荧光探针法检测细胞内活性氧（reactive oxygen species，ROS）水平；采用荧光探针法检测细胞内的Fe2+水平；qRT⁃PCR检测铁死亡相关基因（GPX4、COX2、FTH1和SLC7A11）的表达水平。结果 各浓度小檗碱均可抑制DU145和PC⁃3细胞增殖活力（均P<0.05），且呈浓度依赖性，IC50分别为19.94 μmol/L和20.18 μmol/L。铁死亡抑制剂Ferrostatin1（Fer⁃1）可逆转小檗碱（20 μmol/L）对DU145和PC⁃3细胞增殖活力的抑制作用（均P<0.001），但细胞凋亡和焦亡抑制剂不能恢复细胞活力。20 μmol/L小檗碱可明显降低DU145和PC⁃3细胞内GSH水平和胱氨酸水平（均P<0.001），上调脂质ROS水平和Fe2+水平（均P<0.001），但Fer⁃1可逆转小檗碱诱导的细胞内脂质ROS水平和Fe2+水平（均P<0.001），并恢复GSH和胱氨酸水平（均P<0.001）。此外，小檗碱显著下调DU145和PC⁃3细胞中SLC7A11和GPX4的表达水平（均P<0.05），并上调COX2和FTH1的表达水平（均P<0.01）。过表达SLC7A11可逆转小檗碱对前列腺癌细胞铁死亡的促进作用（均P<0.001）。 结论 小檗碱可能通过抑制SLC7A11表达促进前列腺癌细胞内脂质ROS累积和Fe2+水平上调，进而导致细胞发生铁死亡。

关键词:  , 前列腺癌, 小檗碱, 铁死亡, 脂质氧化

Abstract: Objective To investigate the effect of berberine on ferroptosis in prostate cancer cells and its mechanism. Methods Prostate cancer DU145 and PC⁃3 cells were cultured in vitro, and the effects of berberine with different concentrations (1.560, 3.125, 6.250, 12.500, 25.000, 50.000, 100.000 μmol/L) on the proliferation ability of prostate cancer cells were evaluated by CCK⁃8 assay, and the half⁃inhibitory concentration (IC50) were calculated. The levels of glutathione (GSH), cysteine, reactive oxygen species (ROS), and Fe2+ in both DU145 and PC⁃3 cells were determined by GSH detection assay kit, cysteine assay kit, DCFH⁃DA fluorescent probe assay, and the fluorescent probe, respectively. qRT⁃PCR was performed to detect the expression levels of ferroptosis⁃related genes (GPX4, COX2, FTH1 and SLC7A11). Results Berberine at various concentrations inhibited the proliferation abilities of DU145 and PC⁃3 cells in a concentration⁃dependent way (all P<0.05) , with IC50 values of 19.94 μmol/L and 20.18 μmol/L, respectively. The ferroptosis inhibitor Ferrostatin1 (Fer⁃1) reversed the inhibitory effect of berberine (20 μmol/L) on cell proliferation in DU145 and PC⁃3 cells (all P<0.001), whereas cell apoptosis and pyroptosis inhibitors could not restore cell viability. Treatment with berberine at the concentration of 20 μmol/L significantly decreased the levels of GSH and cysteine in DU145 and PC⁃3 cells (all P<0.001), and increased the levels of lipid ROS and Fe2+ (all P<0.001). However, the Fer⁃1 could reverse berberine⁃induced intracellular lipid ROS level and Fe2+ level (all P<0.001), and restored the levels of GSH and cysteine (all P<0.001). In addition, berberine significantly down⁃regulated the expression levels of SLC7A11 and GPX4 in DU145 and PC⁃3 cells (all P<0.05), while up⁃regulated the expression levels of COX2 and FTH1 (all P<0.01). Overexpression of SLC7A11 reversed the promotive effect of berberine on ferroptosis in prostate cancer cells (all P<0.001). Conclusions Berberine promotes intracellular lipid ROS accumulation and up⁃regulation of Fe2+ levels in prostate cancer cells by inhibiting SLC7A11 expression to induce ferroptosis.

Key words: Prostate cancer, Berberine, Ferroptosis, Lipid oxidation

中图分类号: 

  • R737.25