美洲大蠊提取物对人肝癌细胞Bel-7402作用机制的研究

Effects of Periplaneta Americana L. Extract on Human Hepatoma Cells Bel-7402

关键词:  美洲大蠊提取物  人肝癌细胞Bel-7402  凋亡  线粒体途径

Key words:  Periplaneta americana L. extract  human hepatoma cells Bel-7402  apoptosis  mitochondrial pathway

摘要:

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28.2 μg·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。

Abstract:

OBJECTIVE To investigate the effects of Periplaneta americana L. extract on human hepatoma cells Bel-7402. METHODS The proliferation of human hepatoma cells Bel-7402 was observed by MTT colorimetric, apoptosis was studied by Annexin V-FITC/PI staining. The changes of mitochondrial membrane potential were monitored by flow cytometry, cell apoptosis was detected by DNA ladder experiments, and the protein expressions of Bax, Bcl-2, Caspase-9, Caspase-3, Caspase-8 were tested by western blotting test. RESULTS Periplaneta americana L. extract inhibited the proliferation of human hepatoma cells Bel-7402, IC₅₀ was 28.2 μg·mL^-1. It also induced the apoptosis of human hepatoma cells Bel-7402 and reduced the mitochondrial membrane potential. DNA ladder experiments showed trapezoidal electrophoretogram. Western blotting test showed that the protein expressions of Bax, caspase-9, caspase-3 increased, Bcl-2 decreased, and Caspase-8 expression was not significantly changed. CONCLUSION Periplaneta americana L. extract induces the apoptosis of human hepatoma cells Bel-7402 through mitochondrial pathway.