白藜芦醇诱导鼻咽癌细胞CNE-2Z凋亡的线粒体机制(英文)  

Mitochondrial mechanism of apoptosis induced by resveratrol in nasopharyngeal carcinoma cells CNE-2Z

作　　者：唐旭东[1] 周克元[1] 侯敢[1] 沈晓鸣[1] 

机构地区：[1]广东医学院生物化学与分子生物学研究所,广东湛江524023

出　　处：《中国药理学与毒理学杂志》2004年第3期171-177,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：广东省中医药局重点资助项目 ( 2 0 0 0 0 2 );湛江市科委资助项目 ( 2 0 0 10 6 )~~

摘　　要：目的　探讨白藜芦醇诱导鼻咽癌细胞CNE 2Z凋亡的线粒体机制。方法　用 10 0 μmol·L- 1白藜芦醇分别处理细胞 0 (对照 ) ,2 ,4 ,8,12 ,2 4h和分别用 0 ,2 5 ,5 0 ,10 0 ,2 0 0 μmol·L- 1白藜芦醇处理CNE 2Z细胞 8h ,采用Western印迹分析Bcl 2 ,Bax和胞浆中细胞色素C(CytC)的蛋白水平变化 ,罗丹明 12 3荧光染色后经流式细胞术检测线粒体膜电位(△ψm)变化 ,比色法测定半胱天冬酶 9的活性改变。结果　① 10 0 μmol·L- 1白藜芦醇处理细胞不同时间 ,Bcl 2蛋白表达和△ψm 减少、胞浆中的CytC和半胱天冬酶 9活性增高均呈时间依赖性 (P <0 .0 1) ,但Bax蛋白表达无改变。除Bax以外的其他指标均自白藜芦醇处理 2h即有变化。Bcl 2的蛋白表达受抑和△ψm 的丧失在 4～ 8h间最明显 (与对照组比较P <0 .0 1) ,在 2 4h时已无意义。胞浆中CytC水平和半胱天冬酶 9活性在 8h达高峰 (分别为 0h的 3.0 ,5 .4倍 ) ,2 4h时仍明显高于对照组(P <0 .0 1)。②细胞经不同浓度的白藜芦醇处理 8h后 ,Bcl 2的蛋白表达受抑、△ψm 的丧失、CytC的释放和半胱天冬酶 9活性的升高均具有剂量依赖性 (P <0 .0 1) ,但Bax的蛋白表达未受影响。结论白藜芦醇可能经一个线粒体 /半胱天冬酶 9的特定途径诱导CNE 2Z细胞凋亡 ,但此凋亡?AIM To study whether resveratrol induced apoptosis in nasopharyngeal carcinoma cells CNE 2Z is via a mitochondrial pathway. METHODS CNE 2Z cells were treated with 100 μmol·L-1 resveratrol for 0(control), 2, 4, 8, 12, and 24 h and treated with 0(control), 25, 50, 100, and 200 μmol·L-1 resveratrol for 8 h. The Bcl 2, Bax and cytosolic cytochrome C protein levels were analyzed by Western blot. The mitochondrial membrane potential (△ψm) was detected by flow cytometry following rhodamine 123 fluorescence staining. The caspase 9 activity was determined by colorimetric assay. RESULTS ① After CNE 2Z cells were treated with 100 μmol·L-1 resveratrol for 0(control), 2, 4, 8, 12, and 24 h, respectively, Bcl 2 protein expression and △ψm were decreased, while cytosolic cytochrome C level and caspase 9 activity were increased in a time dependent manner (n=3, P<0.01). But Bax protein expression had no change. All except Bax protein expression had been altered since 2 h. The inhibition of Bcl 2 protein expression and the loss of △ψm were most obvious during 4-8 h (compared with control group P<0.01), but no significance at 24 h. Cytosolic cytochrome C level and caspase 9 activity reached the peak at 8 h and were still much higher than those in control group at 24 h (P<0.01). Cytosolic cytochrome C level and caspase 9 activity were about 3.0 fold and 5.4 fold of those at 0 h, respectively. ②After CNE 2Z cells were treated with 0(control), 25, 50, 100, and 200 μmol·L-1 resveratrol Received date: 2003 08 12 Accepted date: 2003 12 17 Foundation item: The project supported by a key grant from Guangdong Bureau of Traditional Chinese Medicine (200002); and by a grant from Science and Technology Committee of Zhanjiang (200106) Biographies: TANG Xu Dong(1968-), female, native of Fushun, Sichuan Province, associate professor, Master of Medicine, main research fields are biochemical pharmacology and molecular biology of antitumor drugs; ZHOU Ke Yuan(1954-), male, native of Wuchuan, G

关 键 词：白藜芦醇 线粒体 细胞色素C 蛋白BCL-2 蛋白BAX 半胱天冬酶类 

分 类 号：R739.63[医药卫生—肿瘤]