二十碳五烯酸(EPA)体外诱导FRH-0201细胞凋亡机理的初步研究

The Study of the Apoptosis Induced by EPA on Human Cholangiocarcinoma Cell Lines FRH-0201 in Vitro.

目的:观察EPA体外诱导FRH-0201细胞凋亡机理的初步研究。方法:在处于指数生长期的人肝胆管癌细胞株FRH-0201培养剂中添加二十碳五烯酸(EPA),采用流式细胞仪对FRH-0201细胞线粒体跨膜电位的影响、细胞色素c免疫组化染色及Western Bolt检测Casepase-9和Casepase-3的表达。结果:FRH-0201细胞线粒体Δψm下降随EPA剂量的增大而显著增多(P<0.05),EPA处理组细胞内棕色颗粒的强弱和阳性细胞的比例明显高于对照组,差异具有统计学意义(P<0.05,P<0.001),EPA不同浓度处理FRH-0201细胞48h后,Caspase-3和Caspase-9蛋白表达显著提高。结论:EPA可能通过降低线粒体膜电位,释放细胞色素C,激活Caspase-9,从而引发Caspase-3的级联反应,诱导FRH-0201细胞凋亡。

Objective ：To study the apoptosis induced by EPA on Human cholangiocarcinoma cell lines FRH O201 in vitro.Methods ：Exponentially growing Human cholangiocarcinoma cell lines FRH -0201 were exposed to EPA in vitro and a sery of lab test indexes were detected, such as mitochondrial membrane potenial, immunohistochemical staining of cytochrome c, the expression of Caspase -9 and Caspase -3 detected by Western Bolt. Resuhs：After adding EPA, the mitochondrial membrane potential （△ψm） of FRH -0201 cell declined significantly with the increasing doses of EPA （P 〈 0.05）. The intensity of intra cellular brown particle and the proportion of positive cells after adding EPA wele significantly higher than those in the control group, the difference was statistically significant （P 〈0. 05, P 〈0. 001 ）. After adding EPA with different concentration of FRH -0201 cell for 48 hours, the expression of caspase -3 and caspase -9 was significantly increased. Conclusion：EPA can induce the apoptosis of FRH-0201 cells in vitro, it shows that the mechanism may be due to reducing the mitochondrial membrane potential, releasing cytochrome C, activating the cascade reaction induced by caspase - 9, causing caspase 3.