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目的研究没食子酸乙酯(ethyl gallate,EG)对人结肠癌细胞Lovo凋亡的影响并探讨相关机制。方法以体外培养的Lovo细胞作为研究对象,采用倒置显微镜观察细胞生长的变化、Annexin V-FITC/PI双染法和DNA Ladder琼脂糖凝胶电泳法检测没食子酸乙酯的凋亡诱导作用;采用免疫细胞化学法检测cleaved caspase-3,cleaved caspase-9蛋白的表达。结果没食子酸乙酯可明显抑制Lovo细胞生长,倒置显微镜下观察到细胞密度明显降低:各用药组的细胞凋亡率随着药物浓度升高而升高:DNA琼脂糖凝胶电泳可见明显DNA梯状条带:各用药组细胞cleaved caspase-3,cleaved caspase-9基因蛋白阳性表达率明显升高。结论没食子酸乙酯可通过上调cleaved caspase-3和cleaved caspase-9蛋白的表达,激活线粒体通路从而促进Lovo细胞凋亡,这是没食子酸乙酯发挥其抗癌作用的重要机制之一。
Objective To investigate the effect of ethyl gallate (EG) on the apoptosis of human colorectal cancer cell line Lovo and to explore the related mechanisms. Methods Lovo cells cultured in vitro were used as experimental materials. The growth of the cells was observed by inverted microscope. The apoptosis induced by ethyl gallate was detected by Annexin V-FITC / PI double staining and DNA Ladder agarose gel electrophoresis. Immunocytochemistry was used to detect the expression of cleaved caspase-3 and cleaved caspase-9. Results Ethyl gallate significantly inhibited the growth of Lovo cells. The cell density was significantly reduced under inverted microscope. The apoptotic rate of each drug group increased with the increase of drug concentration. DNA ladder agarose gel electrophoresis showed obvious DNA ladder Like bands: The positive expression rates of cleaved caspase-3 and cleaved caspase-9 in each drug group were significantly increased. Conclusion Ethyl gallate can promote the apoptosis of Lovo cells by up-regulating the expressions of cleaved caspase-3 and cleaved caspase-9, activating the mitochondrial pathway, which is one of the important mechanisms by which ethyl gallate exerts its anticancer effect.