白藜芦醇诱导大肠癌SW480细胞发生铁死亡及机制研究

    Resveratrol induces ferroptosis in colorectal cancer SW480 cells and its mechanism

    • 摘要:
      目的研究白藜芦醇对大肠癌SW480细胞铁死亡的激活,并探讨其可能的作用机制。
      方法利用不同剂量白藜芦醇(0、10、20、40、80、160 μmol/L)干预SW480细胞24、48、72 h后,CCK8法分析白藜芦醇对细胞增殖抑制率的影响;不同剂量白藜芦醇(0、20、40、80 μmol/L)干预SW480细胞48 h后,采用蛋白质印迹法分析谷胱甘肽过氧化物酶4(GPX4)、胱氨酸-谷氨酸反向转运体亚基xCT蛋白及二价金属离子转运体1(DMT1)的表达水平,流式细胞仪分析细胞内荧光强度用于监测活性氧(ROS)含量的变化。
      结果白藜芦醇处理SW480细胞后,细胞增殖抑制率明显升高,表现出时间和浓度依赖性(P < 0.05);白藜芦醇能够显著抑制细胞内xCT、GPX4蛋白表达,促使DMT1蛋白表达(P < 0.05);还能诱导SW480细胞内ROS的积累(P < 0.05)。
      结论白藜芦醇通过抑制xCT、GPX4蛋白表达使SW480细胞中ROS大量累积,细胞发生铁死亡,DMT1的激活可能是其机制之一。

       

      Abstract:
      ObjectiveTo study the activation of ferroptosis by resveratrol in colorectal cancer SW480 cells, and to explore its possible mechanism.
      MethodsAfter intervening SW480 cells with different doses of resveratrol(0, 10, 20, 40, 80, 160 μmol/L) for 24, 48 and 72 h, the effect of resveratrol on cell proliferation inhibition rate was analyzed by CCK8 method.After intervening SW480 cells with different doses of resveratrol(0, 20, 40, 80 μmol/L) for 48 h, Western blotting was used to analyze the expression levels of cystine/glutamate antiporter system Xc(xCT), glutathione peroxidase 4(GPX4) and divalent metal transporter 1(DMT1), and flow cytometry was applied to detect the change of reactive oxygen specie(ROS) level which was indicated by fluorescence intensity.
      ResultsAfter SW480 cells were treated with resveratrol, the inhibition rate of cell proliferation was significantly increased, which showed a time- and concentr ation-dependent manner(P < 0.05);the expression level of xCT and GPX4 protein were significantly decreased, and the expression level of DMT1 was greatly increased(P < 0.05);resveratrol also induced the accumulation of ROS in SW480 cells(P < 0.05).
      ConclusionsResveratrol increases the accumulation of ROS by inhibiting the expression of GPX4 and xCT protein to induce ferroptosis in SW480 cells, which may involve the activation of DMT1 expression.

       

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