镰形棘豆对SMMC-7721细胞线粒体膜电位和凋亡相关蛋白表达的影响

目的?研究藏药镰形棘豆对SMMC-7721人肝癌细胞活性、线粒体膜电位，细胞色素C(Cyt C)及Caspase-3蛋白表达的影响与细胞凋亡的关系。方法?以镰形棘豆总黄酮0.05、0.075、0.1g/L处理肝癌细胞SMMC-7721，24h后观察细胞的生长状况；镰形棘豆总黄酮作用于SMMC-7721细胞后，流式细胞仪检测细胞总体凋亡率和线粒体膜电位，Western blot检测对凋亡相关蛋白Cyt C及Caspase-3表达的影响。结果?结果表明，镰形棘豆总黄酮抑制肝癌细胞SMMC-7721的生长，呈浓度依赖关系；镰形棘豆作用后可观察到凋亡细胞的比例升高；可导致细胞的线粒体跨膜电位明显降低，与药物浓度呈负相关。中、高剂量组的细胞中Cyt C及Caspase-3蛋白表达明显升高(P<0.01)。结论?镰形棘豆总黄酮诱导SMMC-7721细胞的凋亡可能是通过线粒体途径，并且与影响Cyt C及Caspase-3的表达有关。为镰形棘豆抗肿瘤的作用机制的实验和临床研究提供依据。 

Effect of Tibetan Medicine Oxytropis Falcata on Mitochondrial Transmembrane Potential and Expression of Apoptosis Related Proteins in SMMC-7721

OBJECTIVE?To explore the influence of Oxytropis falcata on proliferation， mitochondrial membrane potential and expression of cytochrome C (Cyt C) and Caspase-3 proteins in SMMC-7721 cells. METHODS?SMMC-7721 cells were treated with TFOF for 24h. Vitality of SMMC-7721 cells was tested by MTT assay. Flow cytometric measurement was used for investigating the effect of TFOF on induction of apoptosis in SMMC-7721 cells and evaluating the effect on mitochondrial membrane potential in cells. Western blot was applied to investigate changes of Cyt C and Caspase-3 expression in cells. RESULTS?The results showed that TFOF exhibited antiproliferative activity on SMMC-7721 cells in a dose-dependent manner. Flow cytometric analysis demonstrated that TFOF caused a dose-dependent apoptosis of SMMC-7721 cells. After being treated with TFOF of 0.05， 0.075， 0.1g/L， mitochondrial membrane potential levels of SMMC-7721 cells were lower than control group significantly. Western blot showed that Cyt C and Caspase-3 expression of SMMC-7721 cells was up regulated after TFOF treatment (P<0.05~0.01). CONCLUSION?The results suggest that O. falcata may be a pro-apoptotic agent for hepatic cancer cells through mitochondrial apoptosis pathway and via its modulation of Cyt C and Caspase-3 expression.