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目的 :探讨绞股蓝总皂甙 (Gypenosides ,GPS)对人肝癌细胞程序性死亡的影响及机制。方法 :采用MTT法和微量琼脂糖培养克隆形成法检测GPS对体外培养人肝癌细胞株 (HepG2 -A16)细胞生长增殖和克隆形成能力的抑制作用。应用DNA断片法和TUNEL法观测GPS对HepG2-A16细胞程序性死亡的影响 ;同时使用S -P免疫组化法观察GPS对HepG2 -A16细胞Bcl- 2蛋白表达的影响。结果 :1.GPS( 5 0 - 2 5 0mg/L)对HepG2 -A16细胞增殖和克隆形成能力有明显抑制作用(P <0 .0 5 - 0 .0 0 1) ,呈剂量相关性 (P <0 0千 )。 2 .GPS( 5 0 - 2 5 0mg/L)处理HepG2 -A16细胞 2 4小时后 ,DNA琼脂糖电泳表现出细胞程序性死亡特征性的梯形条带图谱 ;TUNEL法染色细胞呈现典型凋亡特征性蓝紫色颗粒。 3.S -P免疫组化法检测HepG2 -A16细胞Bcl - 2蛋白呈阳性染色 ;GPS( 5 0mg/L)孵育 2 4小时后Bcl- 2蛋白呈阴性染色。结论 :GPS具有促进人肝癌细胞程序性死亡作用 ;GPS诱导人肝癌细胞Bcl- 2基因表达下调可能是其促进细胞程序行死亡的主要机制之一。
Objective: To investigate the effect of Gypenosides (GPS) on programmed cell death in human hepatoma cells and its mechanism. METHODS: The inhibition of growth, proliferation and colony-forming ability of human hepatocellular carcinoma cell line (HepG2-A16) cells cultured in vitro by GPS was detected by MTT assay and micro-agar incubation colony formation assay. The DNA fragmentation method and TUNEL method were used to observe the effect of GPS on programmed cell death in HepG2-A16 cells. S-P immunohistochemistry was used to observe the effect of GPS on the expression of Bcl-2 protein in HepG2-A16 cells. Results: 1. GPS (50-250 mg/L) significantly inhibited the proliferation and colony forming ability of HepG2-A16 cells (P < 0.05), showing a dose-related relationship (P < 0.05). <0 0 thousand). 2. GPS (50-250 mg/L) treatment of HepG2-A16 cells for 24 hours, DNA agarose electrophoresis showed a characteristic trapezoidal band pattern of programmed cell death; TUNEL staining of cells showed typical apoptotic characteristics. Blue purple particles. 3. S-P immunohistochemistry method was used to detect the positive expression of Bcl-2 protein in HepG2-A16 cells, and Bcl-2 protein was negatively stained by GPS (50 mg/L) for 24 hours. Conclusion : GPS can promote the programmed death of human hepatocellular carcinoma cells; GPS-induced down-regulation of Bcl-2 gene expression in human hepatocellular carcinoma cells may be one of the mechanisms that promote apoptosis of human hepatoma cells.