细胞周期蛋白依赖性激酶7抑制剂THZ1对脑胶质瘤干细胞干性的调控及机制  

Regulation of THZ1,an inhibitor of cyclin-dependent kinase 7,on stemness of glioma stem cells and its mechanism

作　　者：虎恩喜 何文莹 陶翔 杜沛静 王立斌[1,3] Hu Enxi;He Wenying;Tao Xiang;Du Peijing;Wang Libin(First Clinical Medical College of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Basic Medicine College,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Union Shenzhen Hospital,Huazhong University of Science and Technology,Shenzhen 518000,Guangdong Province,China)

机构地区：[1]宁夏医科大学第一临床医学院,宁夏回族自治区银川市750004 [2]宁夏医科大学基础医学院,宁夏回族自治区银川市750004 [3]华中科技大学协和深圳医院,广东省深圳市518000

出　　处：《中国组织工程研究》2025年第25期5374-5381,共8页Chinese Journal of Tissue Engineering Research

基　　金：深圳市科创委基础研究面上项目(深圳自然科学基金)(JCYJ20230807115807015),项目负责人:王立斌;国家自然科学基金项目(82260610),项目负责人:王立斌;宁夏医科大学校级教育教学改革研究重点项目(NYJY2022011),项目负责人:王立斌。

摘　　要：背景:THZ1是一种细胞周期蛋白依赖性激酶7的抑制剂,可以抑制多种肿瘤细胞的增殖,但THZ1是否可以通过Wnt/β-catenin信号通路影响脑胶质瘤干细胞的干性尚不清楚。目的:探究THZ1对脑胶质瘤细胞U87干性的影响及机制。方法:培养脑胶质瘤细胞U87形成干细胞球并通过Western blot验证干性相关蛋白的表达;采用CCK-8法检测THZ1作用于U87细胞的半数抑制浓度(IC_(50));通过克隆实验、划痕实验、Transwell迁移实验确定THZ1对U87细胞增殖、迁移的影响;分析THZ1处理对U87干细胞成球率及干细胞球体大小的影响;Western blot检测干性相关蛋白CD133、ABCG2、Nanog、OCT4、SOX2和上皮-间充质转化相关蛋白E-cadherin、N-cadherin、Occludin、Snail以及Wnt/β-catenin通路相关蛋白Axin1、β-Catenin、Wnt-5a、GSK3β、Cyclind-1、C-myc的表达变化。结果与结论:①与贴壁细胞相比,U87干细胞干性相关蛋白Nestin、CD133、ABCG2、Nanog、OCT4、SOX2表达明显升高;②与对照组相比,THZ1减弱了U87细胞的增殖和迁移能力;③THZ1抑制U87干细胞成球率及球体大小,下调干性相关蛋白的表达;④THZ1处理后,U87干细胞中N-cadherin、Snail蛋白表达降低,而E-cadherin、Occludin蛋白表达升高;(5)THZ1处理使U87干细胞中Wnt/β-catenin通路相关蛋白Axin1、β-Catenin、Wnt-5a、GSK3β、Cyclind-1、C-myc表达降低。结果表明:THZ1通过下调Wnt/β-catenin信号通路相关分子表达,抑制脑胶质瘤细胞U87的增殖、迁移,抑制U87干细胞成球能力、干性相关蛋白表达和上皮-间充质转化能力。BACKGROUND:THZ1,an inhibitor of cyclin-dependent kinase 7,has been shown to inhibit the proliferation of a variety of tumor cells,but whether THZ1 can affect the stemness of glioma stem cells through the Wnt/β-catenin signaling pathway remains unclear.OBJECTIVE:To investigate the effect of THZ1 on stemness of glioma cell U87 and its mechanism.METHODS:U87 adherent cells were cultured to form stem cell mammospheres.The expressions of stemness related proteins were verified by western blot assay.The effect of THZ1 on half maximal inhibitory concentration(IC_(50))of U87 cells was determined by Cell Counting Kit-8(CCK-8)assays.The effects of THZ1 on proliferation and migration of U87 cells were determined by cell colony-formation assays,cell wound healing assays,and Transwell migration assays.The effect of THZ1 treatment on mammosphere forming rate and mammosphere size of U87 stem cells was analyzed.Stemness associated proteins CD133,ABCG2,Nanog,OCT4,SOX2,epithelial-mesenchymal transformation-related proteins E-cadherin,N-cadherin,Occludin,Snail,and Wnt/β-catenin pathway associated proteins Axin1,β-Catenin,WNT-5A,GSK3β,Cyclind-1,and C-myc were measured by western blot assay.RESULTS AND CONCLUSION:(1)Compared with adherent cells,the expressions of stemness related proteins Nestin,CD133,ABCG2,Nanog,OCT4,and SOX2 were significantly increased.(2)Compared with the control group,THZ1 decreased the proliferation and migration of U87 cells.(3)THZ1 inhibited the mammosphere forming rate and mammosphere size of U87 stem cells.(4)After THZ1 treatment,the expression of N-cadherin and Snail decreased,while the protein expression of E-cadherin and Occludin increased.(5)THZ1 treatment decreased the expression of Wnt/β-catenin pathway related proteins Axin1,β-Catenin,Wnt-5A,GSK3β,Cyclind-1,and C-myc in U87 stem cells.It is concluded that THZ1 can suppress the proliferation and migration of U87 cells,and inhibit the mammosphere forming ability,stemness related protein expression,and epithelial-mesenchymal transformation ability

关 键 词：THZ1 脑胶质瘤 肿瘤干细胞 成球实验 干性标志物 WNT/Β-CATENIN信号通路 上皮-间充质转化 增殖 迁移 

分 类 号：R453.9[医药卫生—治疗学] R394.2[医药卫生—临床医学] R318