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

摘要

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。%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.RE-SULTS 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 he-patic cancer cells through mitochondrial apoptosis pathway and via its modulation of Cyt C and Caspase-3 expression.