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。
Abstract
OBJECTIVE To study the effect of ethyl gallate in inducing apoptosis of human colon carcinoma Lovo cells. METHODS Cultured human colon carcinoma Lovo cells in vitro were used as the research object. Morphological changes of apoptotic cells were observed by inverted microscope.AnnexinV-FITC/PI assay and DNA Ladder assay were performed to study the apoptotic effects. Immunocytochemistry was used to detect the expression of cleaved caspase-9 and cleaved caspase-3 proteins. RESULTS Ethyl gallate had growth inhibition on colon cancer Lovo cells,the density was decrease: the apoptotic rates were increased with dose increase: Agarose gel electrophoresis showed evident DNA fragmentation: the expression of cleaved caspase-9 and cleaved caspase-3 proteins was increased in treated groups. CONCLUSION Ethyl gallate could induce the apoptosis in Lovo cells through up-regulating the expression of cleaved caspase-9 and cleaved caspase-3 protein,and then activated the mitochondrial pathway.
关键词
没食子酸乙酯 /
凋亡 /
线粒体通路 /
人结肠癌细胞Lovo
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Key words
ethyl gallate /
apoptosis /
mitochodrial pathway /
human colon carcinoma Lovo cells
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中图分类号:
R965
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脚注
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基金
国家自然科学基金资助项目(81374021)
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