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目的探讨美洲大蠊提取物对人肝癌细胞Bel-7402的作用机制。方法 MTT比色法观察美洲大蠊提取物对人肝癌细胞Bel-7402增殖的影响,AnnexinV-FITC/PI双染色法研究美洲大蠊提取物对人肝癌细胞Bel-7402凋亡的影响,流式细胞术检测线粒体膜电位,DNA Ladder实验检测细胞凋亡,蛋白印迹法检测Bax、Bcl-2、Caspase-9、Caspase-3、Caspase-8蛋白的表达。结果美洲大蠊提取物可抑制人肝癌细胞Bel-7402增殖,IC50为28.2μg.mL 1,并可诱导人肝癌细胞Bel-7402凋亡,降低线粒体膜电位。DNA Ladder实验可见明显的梯形电泳图谱,蛋白印迹法显示Bax、Caspase-9、Caspase-3蛋白表达增强,Bcl-2蛋白表达减弱,Caspase-8蛋白表达无明显变化。结论美洲大蠊提取物可通过线粒体途径诱导人肝癌细胞Bel-7402凋亡。
Objective To investigate the mechanism of action of Periplaneta americana extract on human hepatoma cell Bel-7402. Methods MTT assay was used to observe the effects of extracts of Periplaneta americana on the proliferation of Bel-7402 human hepatocellular carcinoma cell line. Annexin V-FITC / PI double staining was used to study the effects of extracts of Periplaneta americana on the apoptosis of human hepatocellular carcinoma Bel-7402 cells. Mitochondrial membrane potential was detected by cytometry, apoptosis was detected by DNA Ladder assay, and protein expression of Bax, Bcl-2, Caspase-9, Caspase-3 and Caspase-8 were detected by Western blotting. Results Periplaneta americana extract could inhibit the proliferation of Bel-7402 human hepatocellular carcinoma cells with the IC50 of 28.2μg.mL 1 and induce the apoptosis of Bel-7402 human hepatocellular carcinoma cells and reduce the mitochondrial membrane potential. The DNA ladder showed obvious ladder electrophoresis. Western blotting showed that the expression of Bax, Caspase-9 and Caspase-3 protein increased, the expression of Bcl-2 protein decreased, and the expression of Caspase-8 protein did not change significantly. Conclusion Periplaneta americana extract can induce apoptosis of human hepatocellular carcinoma Bel-7402 through mitochondrial pathway.