Fas、bcl-2和caspase8在去甲斑蝥素诱导食管癌细胞凋亡中的作用及机制
Effect of Fas，bcl-2 and caspase8 on Norcantharidin Induced Esophageal Cancer Cell Line Eca-109 Apotosis and Molecular Mechanism
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摘要: 目的 探讨去甲斑蝥素诱导人食管癌Eca-109细胞凋亡过程中Fas、bcl-2、caspase 8凋亡调节基因的相互关系及可能的作用机制。 方法 流式细胞术分析去甲斑蝥素作用后细胞凋亡及增殖的变化，应用免疫细胞化学技术检测用药前后凋亡相关基因Fas、bcl-2、caspase8表达的变化。 结果 去甲斑蝥素诱导的人食管癌Eca-109细胞发生形态改变。流式细胞仪分析结果发现，食管癌Eca-109细胞在DNA组方图上出现典型的亚二倍体峰即凋亡峰，在细胞周期中的分布也发生了明显的变化。免疫细胞化学结果显示，去甲斑蝥素作用后食管癌Eca-109细胞Fas、caspase8蛋白表达明显升高(P<0.05)，Bcl-2蛋白表达明显降低(P<0.05)。 结论 去甲斑蝥素能抑制食管癌Eca-109细胞增殖并诱导凋亡，其作用可能与上调Fas、caspase8蛋白表达，下调Bcl-2蛋白表达有关。Abstract: Objective To investigate the effect of Fas、bcl-2、caspase 8 on Norcantharidin inhibiting proliferation of Eca-109 cells and inducing its apoptosis and the possible molecule mechanism. Methods The morphology was detected by inverted microscope. The apoptosis and proliferation index of cells were analysed by FCM.The expression of apoptosis associated proteins Fas、bcl-2、caspase8 was measured by immunocytochemical staining. Results The inverted microscope showed that Norcantharidin caused morphological changes of Eca-109 cells. The apoptosis cells were observed on a DNA histogram as subdiploid or pre-G1 peak. The cell cycle distribution changed obviously with treatment of norcantharidin.Immunocytochemical staining showed that the expressions of Fas and caspase 8 protein were induced significantly while the expression of bcl-2 protein was reduced on Eca-109 cells after treated by norcantharidin(P<0.05). Conclusion Norcantharidin may inhibit Eca-109 cell proliferation and induce its apoptosis.The possible mechanism may be related with up-regulating the expressions of Fas、caspase8 and down-regulating the expression of bcl-2 protein.
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