越桔原花青素调控脑胶质瘤细胞生长的机制研究  

Effect of Vaccinium vitis procyanidin on regulation of glioma cell growth

作　　者：钟越[1] 齐玲[1] 沈楠[1] 王玮瑶[1] 田晶[1] 王艳春[1] 

机构地区：[1]吉林医药学院,吉林吉林132013

出　　处：《中国病理生理杂志》2014年第4期603-608,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81201671);吉林省教育厅资助项目(No.2013347);吉林省卫生厅资助项目(No.2013Z203)

摘　　要：目的:探讨越桔原花青素对脑胶质瘤细胞生长的影响。方法:培养脑胶质瘤C6细胞,将其分为对照组以及10、20和40μg/L越桔原花青素组,MTT法检测药物对C6细胞生长的影响,倒置显微镜观察细胞生长的变化,Annexin V/PI染色检测细胞凋亡率的变化,免疫细胞化学法检测Bcl-2与Bax蛋白表达,Western blotting检测Bcl-2、Bax和caspase-3蛋白表达。结果:在24、48和72 h时,10、20和40μg/L越桔原花青素均抑制C6细胞的生长,24和48 h时40μg/L越桔原花青素组、72 h时20和40μg/L越桔原花青素组细胞生长明显受抑制(均P<0.01);倒置显微镜观察发现,各药物组细胞密度随药物浓度升高而减少;Annexin V/PI分析显示,细胞的凋亡率亦随着药物浓度的增加而明显增加;细胞免疫组化结果显示,10、20和40μg/L越桔原花青素组中,C6细胞表达Bcl-2蛋白减少,表达Bax蛋白增加,但均无明显差异;Bax/Bcl-2的比值随着药物浓度升高而增加(P<0.05或P<0.01);Western blotting结果显示,10、20和40μg/L越桔原花青素组中,随着药物浓度升高,细胞表达Bcl-2蛋白减少(P<0.01)而Bax和caspase-3蛋白表达均增加,Bax/Bcl-2的比值也明显增加(P<0.01)。结论:越桔原花青素抑制脑胶质瘤C6细胞的生长,其作用机制与调控Bax蛋白增加和Bcl-2蛋白减少,使Bax/Bcl-2比值增加,从而激活caspase-3,诱导凋亡发生有关。AIM： To explore the effect of Vaccinium viti procyanidin on the growth of glioma cells. METH- ODS ： Glioma C6 cells were cultured and divided into control and 10, 20 and 40 ug/L Vaccinium vitis procyanidin groups. The influence of Vaccinium vitis procyanidin on the growth of C6 cells was measured by MTY assay and the observation un- der inverted microscope. The apoptotic rate was detected by Annexin V/PI staining. The protein expression of Bcl-2 and Bax was determined by immunocytochemistry. The protein levels of Bcl-2, Bax and caspase-3 were also examined by West- ern blotting. RESULTS： The growth of C6 glioma ceils was inhibited by Vaccinium vitis procyanidin at concentrations of 10, 20 and 40 ug/L. The growth was significantly inhibited in 40 ug/L Vaccinium vitis procyanidin group at 24 h and 48 h, and in 20 and 40 ug/L Vaccinium vitis procyanidin groups at 72 h （P 〈0.01 ）. The density of the cells was decreased when the concentration of Vaccinium vitis procyanidin increased. The apoptotic rate was increased when the concentration of Vaccinium vitis procyanidin increased either. The expression of Bcl-2 was decreased and Bax was increased after 10, 20 and 40 ug,/L Vaccinium vitis procyanidin treatments. The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis pro- cyanidin increased （P 〈 0.05 or P 〈 0.01 ）. The expression of Bcl-2 was decreased （P 〈 0.01 ）, and Bax and caspase-3 were increased after 10, 20 and 40 ;~g/L Vaccinium vitis procyanidin treatments. The ratio of Bax/Bcl-2 was increased when the dose of Vaccinium vitis procyanidin increased （P 〈 0.01 ）. CONCLUSION： Vaccinium vitis procyanidin inhibits the growth of glioma cells by down-regulating Bcl-2 protein and up-regulating Bax protein to activate caspase-3, thus indu- cing apoptosis.

关 键 词：越桔原花青素 神经胶质瘤 BAX蛋白 BCL-2蛋白 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R739.4[医药卫生—肿瘤]