SF2523对人源脑胶质瘤干细胞TS576增殖的抑制作用及其机制

doi:10.13481/j.1671-587x.20190615

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (06): 1281-1287.doi: 10.13481/j.1671-587x.20190615

SF2523对人源脑胶质瘤干细胞TS576增殖的抑制作用及其机制

杨智源^1,2, 闻乃妍³, 林杨³, 梁航³, 王乾³, 胡馨丹³, 张灵³, 任辉¹, 郭宝锋¹

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

收稿日期:2019-08-29 出版日期:2019-12-05 发布日期:2019-12-05
通讯作者: 郭宝锋,副主任医师,副教授,硕士研究生导师(Tel:0431-84995114,E-mail:gbf_cc@sohu.com);任辉,主任医师,教授,硕士研究生导师(Tel:0431-84995114,E-mail:hren@jlu.edu.cn) E-mail:gbf_cc@sohu.com;hren@jlu.edu.cn
作者简介:杨智源(1976-),男,吉林省长春市人,副教授,医学硕士,主要从事肿瘤生物学方面的研究。
基金资助:
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）

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

YANG Zhiyuan^1,2, WEN Naiyan³, LIN Yang³, LIANG Hang³, WANG Qian³, HU Xindan³, ZHANG Ling³, REN Hui¹, GUO Baofeng¹

1. Department of General Surgery, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
2. Changchun Medical College, Changchun 130031, China;
3. Department of Pathophysiology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China

Received:2019-08-29 Online:2019-12-05 Published:2019-12-05

摘要/Abstract

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、2.00 μmol·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2 μmol·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2 μmol·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2 μmol·L^-1 SF2523组，利用Annexin Ⅴ/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2 μmol·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2.00 μmol·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，2 μmol·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2 μmol·L^-1 SF2523组G₁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）。Annexin Ⅴ/PI染色检测，作用72 h时，与对照组比较，1和2 μmol·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2 μmol·L^-1SF2523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G₁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。

关键词: SF2523, 脑胶质瘤干细胞, 细胞增殖, 细胞周期, 细胞凋亡

Abstract: 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, 0.50,1.00 2.00 μmol·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 2 μmol·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 2 μmol·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 2 μmol·L^-1 SF2523 groups,and the apoptotic rates of TS576 cells in various groups were detected by Annexin Ⅴ/PI staining. The TS576 cells were divided into control group, 1 and 2 μmol·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 2.00 μmol·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 2 μmol·L^-1 SF2523 group was decreased significantly (P<0.05 or P<0.01). Compared with control group, the percentages of TS576 cells at G₁ phase in 1 and 2 μmol·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 Annexin Ⅴ/PI staining showed that the apoptotic rates of TS576 cells in 1 and 2 μmol·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 μmol·L^-1 SF2523 groups were significantly increased (P<0.01), the expression levels of Bcl-2 and cyclinD1 were significantly decreased (P<0.05 or P<0.01), and the ratio of Bax/bcl-2 was also significantly increased (P<0.01). Conclusion: SF2523 can induce the G₁ phase arrest of TS576 cells by down-regulating the expression of cyclinD1 and promote the apoptosis of TS576 cells by up-regulating the expression of Bax and down-regulating the expression of Bcl-2, and then inhibit the proliferation of TS576 cells.

Key words: SF2523, glioma stem cells, cell proliferation, cell cycle, apoptosis

中图分类号:

R730.264

杨智源, 闻乃妍, 林杨, 梁航, 王乾, 胡馨丹, 张灵, 任辉, 郭宝锋. SF2523对人源脑胶质瘤干细胞TS576增殖的抑制作用及其机制[J]. 吉林大学学报(医学版), 2019, 45(06): 1281-1287.

YANG Zhiyuan, WEN Naiyan, LIN Yang, LIANG Hang, WANG Qian, HU Xindan, ZHANG Ling, REN Hui, GUO Baofeng. Inhibitory effect of SF2523 on proliferation of human glioma stem cells TS576 and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1281-1287.

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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

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