羧胺三唑乳清酸盐抑制小鼠胶质瘤细胞系GL261增殖及线粒体能量代谢

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 957-962.

羧胺三唑乳清酸盐抑制小鼠胶质瘤细胞系GL261增殖及线粒体能量代谢

黄雨晴, 高洪婷, 杨黎星, 陈秋霞, 王钰铖, 鞠瑞^*, 郭磊^*

中国医学科学院基础医学研究所北京协和医学院基础学院药理系,北京 100005

收稿日期:2021-03-10 修回日期:2021-05-19 出版日期:2021-07-05 发布日期:2021-06-17
通讯作者: *leiguo@ibms.cams.cn;jurui1984@163.com
基金资助:
国家自然科学基金(81872897,8200082458);中国医学科学院医学与健康科技创新工程(2016-I2M-1-011)

Carboxyamidotriazole-orotate inhibits the proliferation and mitochondrial energy metabolism of mouse glioma cell line GL261

HUANG Yu-qing, GAO Hong-ting, YANG Li-xing, CHEN Qiu-xia, WANG Yu-cheng, JU Rui^*, GUO Lei^*

Department of Pharmacology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China

Received:2021-03-10 Revised:2021-05-19 Online:2021-07-05 Published:2021-06-17
Contact: *leiguo@ibms.cams.cn;jurui1984@163.com

摘要/Abstract

摘要： 目的研究羧胺三唑乳清酸盐(CTO)对小鼠胶质瘤细胞系GL261增殖和凋亡的影响。方法体外培养小鼠胶质瘤细胞系GL261,实验分为对照组和不同浓度CTO组,采用活细胞计数检测细胞增殖;采用碘化丙啶(PI)染色及流式细胞测量术检测各组GL261的细胞周期;采用annexinⅤ/PI双染色及流式细胞测量术检测各组GL261的细胞凋亡,并计算凋亡率;采用DCFH-DA染色及流式细胞测量术检测各组GL261细胞内活性氧(ROS)的含量;通过Seahorse生物能量仪检测各组GL261细胞的耗氧速率(OCR); 采用三磷酸腺苷(ATP)生物发光法检测各组GL261细胞内ATP的含量。结果与对照组相比,CTO处理组中GL261活细胞数目明显较少,药物作用呈时间-剂量依赖性,20 μmol/L CTO组GL261细胞S期比例明显升高(P＜0.05);与对照组相比,CTO处理组中GL261细胞发生凋亡,药物作用呈时间-剂量依赖性,20 μmol/L CTO组GL261细胞内的ROS含量也明显升高(P＜0.01);与对照组相比,CTO处理组中GL261细胞内的OCR和ATP含量都明显降低(P＜0.01)。结论 CTO可以抑制小鼠胶质瘤细胞系GL261增殖,通过增加ROS的生成促进其凋亡;CTO能够损伤GL261细胞的线粒体呼吸,抑制线粒体中ATP的产生,达到抑制胶质瘤细胞生长的作用。

关键词: 羧胺三唑乳清酸盐, 细胞系GL261, 增殖, 凋亡, 线粒体呼吸

Abstract: Objective To investigate the effects of carboxyamidotriazole-orotate (CTO) on the proliferation and apoptosis of mouse glioma cell line GL261. Methods The proliferation of GL261 cells was detected by live cell counting. Cell cycle of GL261 cells was detected by PI staining and flow cytometry. Apoptosis of GL261 cells was detected by annexinⅤ/PI double staining and flow cytometry; DCFH-DA staining and flow cytometry were used to detect the content of intracellular reactive oxygen species (ROS) of GL261 cells. The oxygen consumption rate (OCR) of GL261 cells was measured in Seahorse bioenergy assay. The ATP content in GL261 cells was detected by ATP detection kit. Results Compared to the control group, the proliferation of GL261 cells in CTO treatment group was significantly inhibited in a time- and dose-dependent way, the proportion of GL261 cells in S phase was significantly increased (P＜0.05) in 20 μmol/L CTO group. Compared with the control group, more apoptosis occurred in GL261 cells of CTO group, and ROS content in GL261 cells of 20 μmol/L CTO group was also significantly increased (P＜0.01). Compared with the control group, the contents of OCR and ATP in GL261 cells was signifi- cantly reduced in the CTO group(P＜0.01). Conclusions CTO inhibits proliferation of GL261 cells and promotes their apoptosis by increased production of ROS. In addition, CTO inhibits the mitochondrial respiration of GL261 cells and the production of ATP in mitochondria, thus inhibits growth of glioma cells.

Key words: carboxyamidotriazole-orotate, cell line GL261, proliferation, apoptosis, mitochondrial respiration

中图分类号:

黄雨晴, 高洪婷, 杨黎星, 陈秋霞, 王钰铖, 鞠瑞, 郭磊. 羧胺三唑乳清酸盐抑制小鼠胶质瘤细胞系GL261增殖及线粒体能量代谢[J]. 基础医学与临床, 2021, 41(7): 957-962.

HUANG Yu-qing, GAO Hong-ting, YANG Li-xing, CHEN Qiu-xia, WANG Yu-cheng, JU Rui, GUO Lei. Carboxyamidotriazole-orotate inhibits the proliferation and mitochondrial energy metabolism of mouse glioma cell line GL261[J]. Basic & Clinical Medicine, 2021, 41(7): 957-962.

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