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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?
Objective To investigate the mitochondrial mechanism of resveratrol-induced apoptosis of nasopharyngeal carcinoma cell line CNE 2Z. Methods The cells were treated with 100 μmol·L-1 resveratrol for 0, 2, 4, 8, 12 and 24 hours and treated with 0, 25, 50, 100, 200 μmol · L-1 resveratrol treatment of CNE 2Z cells 8h, Western blot analysis of Bcl 2, Bax and cytoplasmic cytochrome C (CytC) changes in protein levels, rhodamine 12 3 fluorescence staining by flow cytometry mitochondria Membrane potential (△ ψm) changes, colorimetric assay caspase 9 activity changes. Results ① After resveratrol treatment with 10 0 μmol·L-1 resveratrol, the expression of Bcl-2 protein and △ ψm were decreased at different time points. CytC and caspase 9 activity in cytoplasm increased in a time-dependent manner (P <0 .0 1), but no change in Bax protein expression. Other indicators except Bax resveratrol treatment since 2h that is changed. Bcl 2 protein expression and △ ψm loss between 4 ~ 8h the most obvious (compared with the control group P <0 01), at 24h had no meaning. CytC level and caspase 9 activity in cytoplasm peaked at 8h (3.0 and 5.4 times of 0h respectively), and remained significantly higher at 24h (P <0.01) than that of control group. ② After treated with different concentrations of resveratrol for 8h, the protein expression of Bcl 2, the loss of △ ψm, the release of CytC and the increase of caspase 9 activity were dose-dependent (P <0 .0 1), but the protein expression of Bax was not affected. Conclusion Resveratrol may induce CNE 2Z cell apoptosis via a specific pathway of mitochondrial / caspase 9, but this apoptosis?