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

• 研究论文 • 上一篇    下一篇

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1,高洪婷2,杨黎星1,陈秋霞1,王钰铖2,鞠瑞1,郭磊1   

  1. 1. 中国医学科学院基础医学研究所
    2. 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
  • 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: 郭磊 E-mail:guoleistu@126.com
  • 基金资助:
    国家自然科学基金;中国医学科学院医学与健康科技创新工程

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

  • Received:2021-03-10 Revised:2021-05-19 Online:2021-07-05 Published:2021-06-17
  • Contact: Lei GUO E-mail:guoleistu@126.com

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)染色及流式细胞术检测各组GL261的细胞周期;采用AnnexinⅤ/PI双染色及流式细胞术检测各组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,20μmol/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,20μmol/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 count. 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) in 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 with 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 20μmol/L CTO group. Compared with the control group, more apoptosis occurred in GL261 cells of CTO group, and ROS content in GL261 cells of 20μmol/L CTO group was also significantly increased (P < 0.01). Compared with the control group, the contents of OCR and ATP in GL261 cells were significantly reduced in the CTO group(P < 0.01). Conclusions CTO can inhibit the proliferation of GL261 cells and promote their apoptosis by increasing the production of ROS. In addition, CTO can inhibit the mitochondrial respiration of GL261 cells and inhibit the production of ATP in mitochondria, thus impair the growth of glioma cells.

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

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