SF2523对人源脑胶质瘤干细胞TS576增殖的抑制作用及其机制  

Inhibitory effect of SF2523 on proliferation of human glioma stem cells TS576 and its mechanism

作　　者：杨智源 闻乃妍[3] 林杨 梁航[3] 王乾[3] 胡馨丹[3] 张灵 任辉[1] 郭宝锋[1] YANG Zhiyuan;WEN Naiyan;LIN Yang;LIANG Hang;WANG Qian;HU Xindan;ZHANG Ling;REN Hui;GUO Baofeng(Department of General Surgery,China-Japan Union Hospital,Jilin University,Changchun 130033,China;Changchun Medical College,Changchun 130031,China;Department of Pathophysiology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学中日联谊医院普通外科,吉林长春130033 [2]长春医学高等专科学校,吉林长春130031 [3]吉林大学基础医学院病理生理学系,吉林长春130021

出　　处：《吉林大学学报（医学版）》2019年第6期1281-1287,共7页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81773217);吉林省科技厅国际合作项目资助课题(20190701065GH)

摘　　要：目的:探讨PI3K和BRD4双重抑制剂SF2523对人源脑胶质瘤干细胞TS576增殖的抑制作用,并初步阐明其作用机制。方法:培养人源脑胶质瘤干细胞TS576,将其分为对照组和0.25、0.50、1.00、2.00μmol·L^-1 SF2523组。采用CCK-8法检测各组TS576细胞存活率;将TS576细胞分为对照组和2μmol·L^-1 SF2523组,利用细胞生长计数法检测各组TS576细胞数;将TS576细胞分为对照组和1及2μmol·L^-1 SF2523组,采用流式细胞术检测各组不同细胞周期TS576细胞百分比;将TS576细胞分为对照组和1及2μmol·L^-1 SF2523组,利用AnnexinⅤ/PI染色法检测各组TS576细胞凋亡率;将TS576细胞分为对照组和1及2μmol·L^-1 SF2523组,采用Western blotting法检测各组TS576细胞中B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)和cyclinD1蛋白表达水平。结果:CCK-8检测,作用24、48和72 h时,与对照组比较,0.25、0.50、1.00和2.00μmol·L^-1 SF2523组TS576细胞存活率均明显降低(P<0.01)。细胞生长计数法检测,与对照组比较,2μmol·L^-1 SF2523组细胞数明显减少(P<0.05或P<0.01)。流式细胞术检测,作用72 h时,与对照组比较,1和2μmol·L^-1 SF2523组G 1期TS576细胞百分比升高(P<0.05),S期TS576细胞百分比降低(P<0.05)。AnnexinⅤ/PI染色检测,作用72 h时,与对照组比较,1和2μmol·L^-1 SF2523组细胞凋亡率均明显升高(P<0.01)。Western blotting法检测,作用72 h时,与对照组比较,1和2μmol·L^-1 SF2523组TS576细胞中Bax蛋白表达水平升高(P<0.01),Bcl-2和cyclinD1蛋白表达水平降低(P<0.05或P<0.01),Bax/Bcl-2比值升高(P<0.01)。结论:SF2523通过下调cyclinD1表达引起TS576细胞G 1期阻滞,通过上调Bax表达、下调Bcl-2表达促进TS576细胞凋亡,从而抑制TS576细胞增殖。Objective:To investigate the inhibitory effect of SF2523,a dual inhibitor of BRD4 and PI3K,on the proliferation of human glioma stem cells TS576,and to preliminarily elucidate its mechanism.Methods:The human glioma stem cells TS576 were cultured and divided into control group and 0.25,0.50,1.002.00μmol·L^-1 SF2523 groups,and CCK-8 method was used to detect the survival rates of TS576 cells.The TS576 cells were divided into control group and 2μmol·L^-1 SF2523 group,and the number of TS576 cells in various groups was detected by cell growth counting method.The TS576 cells were divided into control group,1 and 2μmol·L^-1 SF2523 groups,and the percentages of TS576 cells at different cell cycles in various groups were examined by flow cytometry.The TS576 cells were divided into control group,1 and 2μmol·L^-1 SF2523 groups,and the apoptotic rates of TS576 cells in various groups were detected by AnnexinⅤ/PI staining.The TS576 cells were divided into control group,1 and 2μmol·L^-1 SF2523 groups,and the expression levels of cyclinD1,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)proteins in the TS576 cells in various groups were detected by Western blotting method.Results:Compared with control group,the proliferation rates of TS576 cells in 0.25,0.50,1.00 and 2.00μmol·L^-1 SF2523 groups at 24,48 and 72 h after treatment were significantly decreased(P<0.01).Compared with control group,the number of TS576 cells in 2μmol·L^-1 SF2523 group was decreased significantly(P<0.05 or P<0.01).Compared with control group,the percentages of TS576 cells at G 1 phase in 1 and 2μmol·L^-1 SF2523 groups were increased(P<0.05),the percentage of TS576 cells at S phase was decreased(P<0.05).The results of AnnexinⅤ/PI staining showed that the apoptotic rates of TS576 cells in 1 and 2μmol·L^-1 SF2523 groups were significantly increased at 72 h after treatment compared with control group(P<0.01).The results of Western blotting method showed that the expression levels of Bax protein in the TS576 cells in 1 and 2�

关 键 词：SF2523 脑胶质瘤干细胞 细胞增殖 细胞周期 细胞凋亡 

分 类 号：R730.264[医药卫生—肿瘤]