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中国癌症防治杂志

中国癌症防治杂志 ›› 2024, Vol. 16 ›› Issue (2): 158-165.doi: 10.3969/j.issn.1674-5671.2024.02.04

• 基础研究 • 上一篇    下一篇

蟾毒它灵诱导急性早幼粒细胞白血病HL-60细胞铁死亡的机制研究

  

  1. 陕西中医药大学第二临床医学院；陕西中医药大学第二附属医院；陕西中医药大学制药厂
  • 出版日期:2024-04-25 发布日期:2024-05-08
  • 通讯作者: 李宏 E-mail：sxefyy.lh@163.com
  • 基金资助:
    陕西省自然科学基础研究计划项目（2024JC-YBMS-742）；陕西省中医药管理局第二批省级中医药中青年科技骨干人才项目（2023-ZQNG-006）;咸阳市创新服务能力支撑计划项目（L2023-CXNL-CXRC-015）

A study on the mechanism of Bufotalin-induced ferroptosis in acute promyelocytic leukemia HL-60 cells

  • Online:2024-04-25 Published:2024-05-08

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摘要： 目的 探究蟾毒它灵（Bufotalin）对急性早幼粒细胞白血病（acute promyelocytic leukemia，APL）细胞HL⁃60铁死亡的作用及机制。 方法 用不同浓度（0.1、0.5、1.0、2.0、4.0 μg·mL-1）蟾毒它灵分别作用于APL细胞HL⁃60，使用光学显微镜观察细胞的形态学变化；采用CCK⁃8法检测细胞的存活率情况；采用Western blot法检测细胞中铁死亡相关蛋白CD71、xCT、FTH1、GPX4的表达水平；采用谷胱甘肽（GSH/GSSG）检测试剂盒检测细胞内GSH和GSSG的含量变化；采用脂质氧化（MDA）检测试剂盒测定细胞内MDA的含量变化。将0.5 μg·mL-1蟾毒它灵、0.1 μmol·L-1 Fer⁃1和0.5 μg·mL-1蟾毒它灵+0.1 μmol·L-1 Fer⁃1分别作用于HL⁃60细胞，使用光学显微镜观察细胞形态学变化，并用Western blot法检测铁死亡相关蛋白的表达水平。 结果 不同浓度蟾毒它灵处理HL⁃60细胞后，细胞形态呈现出哑铃型、梭形等不规则形状，细胞的存活率降低，且呈时间⁃剂量依赖关系。与对照组相比，蟾毒它灵可降低铁死亡相关蛋白GPX4、xCT、FTH1的表达水平和总谷胱甘肽的含量（均P<0.01），升高CD71蛋白的表达水平和MDA含量（均P<0.05）。蟾毒它灵与铁死亡抑制剂Fer⁃1联合处理H60细胞后，与对照组比较，HL⁃60细胞的生长数量和形态学无明显变化，CD71、xCT、FTH1、GPX4蛋白表达水平的差异也无统计学意义（均P>0.05）。结论 蟾毒它灵可抑制APL细胞HL⁃60的增殖并诱导铁死亡，其可能通过GPX4介导的抗氧化途径和铁代谢途径实现。

关键词: 蟾毒它灵, 急性早幼粒细胞白血病, HL-60细胞, 铁死亡

Abstract: Objective To investigate the effect and mechanism of Bufotalin on ferroptosis in acute promyelocytic leukemia (APL) HL⁃60 cells. Methods APL HL⁃60 cells were treated with Bufotalin at different concentrations (0.1, 0.5, 1.0, 2.0, and 4.0 μg·mL-1, respectively), and the morphological changes of cells were observed using a light microscope. The survival rate of cells was detected by the CCK⁃8 method. The expression levels of ferroptosis⁃related proteins CD71, xCT, FTH1 and GPX4 in HL⁃60 cells were detected by Western blot. The changes of contents of intracellular GSH and GSSG were detected by GSH/GSSG detection kit. The changes of contents of intracellular MDA in HL⁃60 cells were determined by MDA detection kit. HL⁃60 cells were treated with 0.5 μg·mL⁃1 Bufotalin 0.1 μmol·L⁃1 Fer⁃1 and  0.5 μg·mL⁃1 Bufotalint 0.1 μmol·L⁃1 Fer⁃1, respectively, and the morphological changes of cells were observed using a light microscope, the expression levels of ferroptosis⁃related proteins were detected by Western blot. Results After treatment with different concentrations of Bufotalin, HL⁃60 cells showed irregular shapes, such as dumbbell shape and spindle shape, and the survival rate of the cells decreased in a time⁃dose dependent relationship. Compared with the control group, Bufotalin could reduce the expression levels of ferroptosis⁃related proteins GPX4, xCT and FTH1, as well as the content of total glutathione (all P<0.01), and increase the expression level of CD71 protein and the content of MDA (all P<0.05). Compared with the control group, the growth number and morphology of HL⁃60 cells were not significantly changed after combined treatment of Bufotalin and Fer⁃1, the protein expression levels of CD71, xCT, FTH1 and GPX4 were not statistically significant (all P>0.05). Conclusions  Bufotalin can inhibit the proliferation of APL HL⁃60 cells and induce ferroptosis, the underlying molecular mechanism is probably realized through GPX4⁃mediated antioxidant pathway and iron metabolism pathway. 

Key words: Bufotalin, Acute promyelocytic leukemia, HL-60 cells, Ferroptosis

中图分类号: 

  • R737.9