胡桃醌对肝癌HepG2细胞的增殖抑制及诱导凋亡作用

目的:探讨胡桃醌对肝癌HepG2细胞增殖的影响,阐明其可能的作用机制。方法:选取处于对数生长期的肝癌细胞株HepG2分为空白对照组和不同剂量(40、80、120、160和200μmol·L1)胡桃醌组。采用MTT法观察胡桃醌对HepG2细胞的抑制作用,并计算出半数抑制浓度(IC50),依据IC50确定胡桃醌的有效浓度;在光学显微镜下观察细胞形态学变化;免疫化学法检测HepG2细胞中Caspase-3蛋白的表达;流式细胞术测定120μmol·L1胡桃醌对HepG2细胞凋亡周期的影响。结果:胡桃醌的IC50为139.8881μmol·L1。与空白对照组比较,各剂量胡桃醌组HepG2细胞增殖抑制率明显升高(P<0.05或P<0.01),并呈剂量依赖性。形态学观察,胡桃醌组细胞体积缩小,连接消失。120μmol·L1胡桃醌组HepG2细胞的生长周期发生变化,出现明显的G2/M期阻滞。免疫细胞化学染色,与空白对照组比较,120μmol·L1胡桃醌组HepG2细胞中Caspase-3蛋白表达明显增加。结论:胡桃醌能诱导HepG2细胞发生凋亡,其机制可能与Caspase通路相关。

Proliferation inhibition and apoptosis induction of Juglone on human hepatoma HepG2 cells

Objective To explore the effect of Juglone on proliferation of liver cancer HepG2 cells and to clarify its possible mechanism.Methods The human hepatoma HepG2 cells were divided into different doses of Juglone(40,80,120,160 and 200 μmol·L-1) groups and control group.The proliferation inhibition of Juglone on HepG2 cells was measured by MTT assay,and IC50 was calculated based on the effective concentration.Optical microscope was used to observe the morphological changes of HepG2 cells.Meanwhile the expression of Caspase-3 protein was analyzed by immunocytochemistry.The cell cycle of HepG2 cells was detected by flow cytometry.Results The IC50 of Juglone was 139.888 1 μmol·L-1.Compared with control group,the inhibitory rates of proliferation of HepG2 cells in different doses of Juglone groups were increased significantly(P<0.05 or P<0.01) in a dose-dependent manner.The morphological observation showed that the cells in different does of Juglone groups were smaller and the connection disappeared.In 120 μmol·L-1 Juglone group the cell cycle of HepG2 cells was changed and the cells were markedly blocked in the G2/M phase.The immunocytochemistry staining results indicated that the Caspase-3 protein expression in 120 μmol·L-1Juglone group was higher than that in control group.Conclusion Juglone could induce the apoptosis of HepG2 cells,and the mechanism may be related to Caspase pathway.