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目的研究死亡受体通路在没食子酸乙酯诱导结肠癌细胞Lovo凋亡过程中的作用。方法以结肠癌细胞Lovo为研究对象,采用AO/EB荧光染色法观察药物作用后凋亡细胞形态学变化;免疫细胞化学法检测用药前后凋亡相关基因Caspase-8,Fas,Survivin蛋白的表达。结果没食子酸乙酯作用后,AO/EB双染后荧光显微镜下观察Lovo细胞呈典型的凋亡形态学改变。免疫细胞化学结果表明,实验组活性Caspase-8蛋白和Fas蛋白阳性表达率增强,而Survivin阳性表达率减弱,与对照组相比具有显著差异。结论没食子酸乙酯可通过上调Fas和活性Caspase-8蛋白的表达,下调Survivin蛋白的表达。死亡受体通路参与没食子酸乙酯诱导Lovo细胞凋亡的过程,这可能是EG发挥抗肿瘤作用机制之一。
Objective To investigate the role of death receptor pathway in the apoptosis of Lovo cells induced by ethyl gallate. Methods Colon cancer cell line Lovo was used as research object. AO / EB staining was used to observe the morphological changes of apoptotic cells. The expression of apoptosis related genes Caspase-8, Fas and Survivin were detected by immunocytochemistry. Results Ethyl gallate, AO / EB double staining under a fluorescence microscope Lovo cells were observed typical apoptotic morphological changes. Immunocytochemistry results showed that the positive expression rate of active Caspase-8 protein and Fas protein in the experimental group increased while the positive expression rate of Survivin decreased, which was significantly different from the control group. Conclusion Ethyl gallate can down-regulate the expression of Survivin protein by up-regulating Fas and Caspase-8 protein expression. The death receptor pathway is involved in the induction of Lovo cell apoptosis by ethyl gallate, which may be one of the mechanisms by which EG exerts antitumor effects.