黄芩素通过调控Wnt/β-catenin信号通路抑制人胶质母细胞瘤细胞MGMT表达  

Baicalein Inhibits MGMT Expression in Glioblastoma by Modulating Wnt/β-catenin Pathway

作　　者：卫翔宇 陈正 沈霖 李世亭[1] 郑学胜[1] WEI Xiang-yu;CHEN Zheng;SHEN Lin;LI Shi-ting;ZHENG Xue-sheng(Department of Neurosurgery, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200092, Chin)

机构地区：[1]上海交通大学医学院附属新华医院神经外科,上海200092

出　　处：《现代生物医学进展》2018年第8期1414-1418,共5页Progress in Modern Biomedicine

基　　金：上海市科学技术委员会科研项目(14DZ1930303)

摘　　要：目的:研究黄芩素在体外对人胶质母细胞瘤T98G细胞和GBM-X细胞MGMT(O6-甲基鸟嘌呤-DNA甲基转移酶)表达的作用以及相关的信号通路机制。方法:将T98G细胞和GBM-X细胞在常氧和缺氧条件下分成对照组与处理组,采用Western Blot(蛋白质印迹法)检测黄芩素作用于两种细胞72 h后MGMT及其相关信号通路蛋白的表达水平,CCK-8试验检测黄芩素、替莫唑胺以及二者联用对两种细胞的毒性作用,探讨联合用药的可能性。结果:T98G细胞常氧/缺氧条件下黄芩素处理组MGMT蛋白表达水平(0.757±0.058,0.714±0.049)较对照组(1.000±0.101,1.000±0.072)显著下降(P<0.05,P<0.01),GBM-X细胞常氧/缺氧条件下黄芩素处理组MGMT蛋白表达水平(0.843±0.027,0.697±0.052)相比对照组(1.000±0.031,1.000±0.082)明显降低(P<0.01,P<0.01)。同对照组相比,T98G、GBM-X细胞黄芩素处理后TCF1/TCF7(T细胞因子)、LEF1(淋巴样增强因子)、Survivin(生存素)蛋白表达水平明显下降,差异有统计学意义。经CCK8试剂检测,T98G、GBM-X细胞药物作用24、48、72 h后,替莫唑胺组与对照组相比无明显统计学差异,黄芩素联合替莫唑胺组较对照组吸光度值显著降低,差异有统计学意义。结论:黄芩素可通过调控Wnt/β-catenin(β-连环蛋白)信号通路稳定抑制人胶质母细胞瘤细胞MGMT蛋白表达,黄芩素联合替莫唑胺可降低MGMT阳性胶质母细胞瘤细胞对替莫唑胺的耐药性,增强替莫唑胺细胞毒作用,发挥替莫唑胺化疗效果。Objective： To investigate the effect of baicalein on the expression of MGMT in human glioblastoma T98G cells and GBM-X cells in vitro and the related signaling pathways. Methods： Divide T98G cells and GBM-X cells into control group and experimental group under normoxia and hypoxia conditions. Then detect the expression of MGMT and related signal channel proteins by Western Blot after 72 h treatment and detect the cytotoxicity of baicalein, temozolomide and combined pharmacotherapy by CCK-8 test to explore the possibility of combination therapy. Results： The expression of T98G cells MGMT protein in the baicalein treatment group（0.757 ±0.058, 0.714 ±0.049） was significantly lower than that of the control group（1.000 ±0.101, 1.000 ±0.072）under normoxia/hypoxia condition（P〈0.05, P〈0.01）. Compared with the control group（1.000 ±0.031, 1.000 ±0.082）, the expression of MGMT protein in GBM-X cells in the experimental group（0.843±0.027, 0.697±0.052）decreased significantly under normoxia/hypoxia condition（P〈0.01,P〈0.01）. Compared with the control group, the expression level of TCF1/TCF7, LEF1 and Survivin protein decreased significantly after baicalein treatment in T98G and GBM-X cells. After CCK8 detection, there was no significant difference between the temozolomide group and the control group after 24, 48 and 72 h in T98G and GBM-X cells, but the value of CCK8 in the combination of baicalein and temozolomide group was significantly lower than that of the control group with the difference statistically significant.Conclusions： Baicalein stably inhibits the expression of MGMT in human glioblastoma cells by regulating the Wnt/β-catenin signaling pathway and reduces temozolomide resistance of MGMT positive glioblastoma cells and then enhances the cytotoxic effect of temozolomide.

关 键 词：黄芩素 T98G细胞 GBM-X细胞 MGMT WNT/Β-CATENIN信号通路 

分 类 号：R-33[医药卫生] R739.41