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目的:探讨重楼皂苷Ⅶ(Paris saponin Ⅶ,PSⅦ)对人结肠癌SW480细胞凋亡和周期的影响及其机制。方法:采用Cell Counting Kit-8(CCk-8)试剂盒方法观察PSⅦ对SW-480细胞增殖的影响;流失细胞术研究PSⅦ对SW480细胞凋亡和周期的作用;培养SW-480细胞并给予1.0μmol/L、2.0μmol/L和4.0μmol/L PSⅦ 24 h后,采用Western blot法检测凋亡相关蛋白Caspase-3、Caspase-8、Caspase-9、Bax和Bcl-2及周期相关蛋白Cdk-4、Cdk-6、Cyclin D1和p21的表达变化。结果:PSⅦ可明显抑制SW-480细胞增殖,并诱导其凋亡,其IC50值为5.25±0.46μmol/L;PSⅦ可促进SW-480细胞Caspase-3、Caspase-8、Caspase-9、Bax及p21的表达,降低Bcl-2、Cdk-4、Cdk-6和Cyclin D1的表达。结论:PSⅦ可以通过线粒体和死亡受体途径诱导SW-480细胞凋亡;将SW-480细胞周期阻滞于G1期是通过上调p21,抑制Cdk-4、Cdk-6与Cyclin D1周期调控蛋白的表达。
Objective: To investigate the effect and mechanism of Paris saponin Ⅶ (PSⅦ) on apoptosis and cycle of human colon cancer SW480 cells. Methods: The effect of PSⅦ on proliferation of SW-480 cells was observed by Cell Counting Kit-8 (CCk-8) kit. Flow cytometry was used to study the effect of PSⅦ on apoptosis and cycle of SW480 cells. SW-480 cells were cultured and treated with 1.0 The apoptosis-related proteins Caspase-3, Caspase-8, Caspase-9, Bax and Bcl-2 and the expression of cyck-related protein Cdk- 4, Cdk-6, Cyclin D1 and p21 expression changes. Results: PSⅦ could significantly inhibit the proliferation and induce apoptosis of SW-480 cells with the IC50 value of 5.25 ± 0.46μmol / L. PSⅦ promoted the expression of Caspase-3, Caspase-8, Caspase-9 and Bax p21 expression, reduce the expression of Bcl-2, Cdk-4, Cdk-6 and Cyclin D1. CONCLUSIONS: PSⅦ can induce apoptosis of SW-480 cells through the mitochondrial and death receptor pathways. Blocking G1-phase of SW-480 cell cycle inhibits the expression of Cdk-4, Cdk-6 and Cyclin D1 expression.