Juglone-induced apoptosis in human gastric cancer SGC-7901 cells via the mitochondrial pathway
Juglone-induced apoptosis in human gastric cancer SGC-7901 cells via the mitochondrial pathway
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胡桃醌通过线粒体途径诱导人胃癌SGC-7901细胞凋亡
DOI：
10.1016/j.etp.2009.09.010
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发表时间：
2011-01-01
影响因子：
--
通讯作者：
Zou, Xiang
中科院分区：
文献类型：
--
作者：
Ji, Yu-Bin;Qu, Zhong-Yuan;Zou, Xiang
本研究旨在探讨胡桃醌对人胃癌SGC-7901细胞凋亡的影响。采用磺酰罗丹明B（SR B）法检测胡桃醌对人胃癌细胞SGC-7901的细胞毒活性。透射电镜观察细胞形态学变化。流式细胞仪检测细胞凋亡率、活性氧水平、线粒体跨膜电位及细胞色素c蛋白表达。Western blot检测Bcl-2和Bax蛋白表达。用酶标仪测定胱天蛋白酶3活性。结果表明：SGC-7901细胞的Gl（50）值为36.51 ± 1.05 μ mol/L（24 h）和25.37 ± 1.19 μ mol/L（48 h）。胡桃醌（5、10、15和20 μ mol/L）作用24 h后，细胞出现典型的凋亡形态学改变，凋亡率呈剂量依赖性增加。用相同剂量的胡桃醌处理细胞24 h后，与对照组相比，ROS水平显著升高，Bcl-2表达显著下调，Bax表达显著上调。线粒体跨膜电位显着降低，细胞色素c蛋白的表达显着高于对照组。Caspase 3以浓度依赖性方式被激活。总之，胡桃醌可以通过线粒体途径诱导SGC-7901细胞凋亡，该途径似乎是由ROS的产生和Bcl-2/Bax比值的降低介导的。Crown版权所有（C）2009由Elsevier GmbH发布。All rights reserved.
This study was designed to investigate the effect of juglone on the apoptosis of human gastric cancer SGC-7901 cells. The cytotoxic activity of juglone on SGC-7901 cells was tested by the sulforhodamine B (SRB) assay. The morphological changes in the cells were observed by transmission electron microscopy (TEM). The apoptotic rate, the level of reactive oxygen species (ROS), mitochondrial transmembrane potential and the expression of cytochrome c protein were detected by flow cytometry (FCM). The expression of Bcl-2 and Bax proteins were examined by Western blot. Caspase 3 activity was determined with a microplate reader. Our results were as follows: the Gl(50) values for SGC-7901 cells were 36.51 +/- 1.05 mu mol/L (24 h) and 25.37 +/- 1.19 mu mol/L (48 h). After 24 h of exposure to juglone (5, 10, 15 and 20 mu mol/L), the cells presented the typical morphological changes of apoptosis, and the rate of apoptosis was found to increase in a dose-dependent manner. After cells were treated with juglone at the same dose for 24h. the level of ROS was significantly higher, the expression of Bcl-2 was significantly down-regulated and the expression of Bax was significantly up-regulated compared to the control. The mitochondrial transmembrane potential was significantly lower, and the expression of the cytochrome c protein was significantly higher relative to the control. Caspase 3 was activated in a concentration-dependent manner. In conclusion, juglone can induce apoptosis in SGC-7901 cells through a mitochondrial pathway that seems to be mediated by the generation of ROS and a reduction in the Bcl-2/Bax ratio. Crown Copyright (C) 2009 Published by Elsevier GmbH. All rights reserved.