槲皮黄酮对膀胱癌细胞抑制作用的机制研究  

Mechanism in growth inhibition of quercetin on human bladder cancer cell line

作　　者：谭大清[1] 刘孝华[1] 

机构地区：[1]湖北民族学院附属民大医院泌尿外科,湖北恩施445000

出　　处：《中国中药杂志》2017年第9期1742-1751,共10页China Journal of Chinese Materia Medica

摘　　要：以BIU-87细胞为模型,探讨槲皮黄酮对膀胱癌细胞的抑制作用机制。体外培养人膀胱癌细胞株BIU-87,将不同浓度槲皮黄酮加入处于对数生长期的BIU-87细胞中共培养,采用CCK-8法检测槲皮黄酮对BIU-87细胞增殖的抑制作用;流式细胞仪检测分析槲皮黄酮诱导BIU-87细胞的凋亡作用和细胞周期;Western blot技术检测分析了细胞中抗凋亡蛋白Bal-2,Bal-x L的表达量及TAK1/JNK信号通路中蛋白的表达水平。CCK-8实验发现,不同浓度槲皮黄酮处理BIU-87细胞24 h或48h后,药物对BIU-87细胞增殖抑制作用均明显高于对照组(P<0.05);流式细胞术结果显示,不同浓度槲皮黄酮对BIU-87细胞的凋亡率均显著高于对照组,呈剂量时间依赖性,细胞周期G_0/G_1期细胞逐渐减少,G_2/M期细胞逐渐增多,S期细胞变化不明显;Western blot检测发现槲皮黄酮处理的BIU-87细胞中Bal-2,Bal-x L蛋白表达水平降低,并且p-TAK1,p-MKK4/7,p-JNK表达水平也降低。槲皮黄酮可抑制BIU-87细胞增殖,促进BIU-87细胞凋亡,其机制可能与抑制TAK1/JNK信号通路进一步降低了Bal-2,Bal-x L的表达有关。To investigate the inhibitory mechanism of quercetin on growth of human bladder cancer cell line（BIU-87）. BIU-87 cells were cultured in vitro,and co-cultured with varying concentrations of quercetin,and the anti-proliferative activity was determined by CCK-8 assay. Apoptosis of quercetin-induced BIU-87 cells and cell cycle were determined by flow cytometry analysis. Expressions of Bal-2 and Bal-x L,and related proteins in TAK1/JNK signal pathway were measured using Western blot analysis. After treatment with quercetin for 24 h and 48 h,the proliferation of BIU-87 cells was significantly suppressed in a dose-dependent manner according to CCK-8 assay（ P〈0. 05）. The flow cytometry analysis indicated that each group of quercetin leads to a significant higher percentage of apoptosis of BIU-87 cells than control group after treatment with quercetin for 24 h and 48 h; In G_0/G_1 period,cells reduced,while the amount of cells in G2/M period increased,and cells in S period remained the same amount. Expressions of Bal-2,Bal-x L,p-TAK1,p-MKK4/7,p-JNK decreased in BIU-87 cells after treatment with quercetin. Quercetin could inhibit the proliferation and promote the apoptosis of BIU-87 cells. The mechanism may be correlated with the inhibition of TAK1/JNK signaling pathway,which led to the further decrease in expressions of Bal-2 and Bal-x L.

关 键 词：槲皮黄酮 膀胱癌细胞 增殖 凋亡 

分 类 号：R285[医药卫生—中药学]