上海交通大学学报(医学版)

• 论著(基础研究) • 上一篇    下一篇

线粒体二羧基载体蛋白SLC25A10对脑胶质瘤细胞增殖的影响及机制

吕依宣 1,王晓娜 1*,吴金亮 2,许艳 1,糜军 1   

  1. 上海交通大学 1.基础医学院生物化学与分子细胞生物学系,上海 200025;2.医学院附属第九人民医院影像科,上海 201999
  • 出版日期:2016-11-28 发布日期:2016-11-29
  • 通讯作者: 许艳,电子信箱:yan_xu@sjtu.edu.cn;糜军,电子信箱:jmei@sjtu.edu.cn。
  • 作者简介:吕依宣(1990—),女,硕士生;电子信箱:lvyixuan1990@qq.com;王晓娜(1987—),女,助理实验师,硕士;电子信箱: iwangxiaona@163.com。*为共同第一作者。
  • 基金资助:

    国家自然科学基金(81602509);上海高校青年教师培养资助计划(ZZjdyx15016)

Effects of mitochondrial dicarboxylate carrier SLC25A10 on the proliferation of glioma cells and relevant mechanisms

Lü Yi-xuan 1, WANG Xiao-na 1*, WU Jin-liang 2, XU Yan 1, MI Jun 1   

  1. 1. Department of Biochemistry and Molecular Cell Biology, Basic Medicine Faculty of Shanghai Jiao Tong University, Shanghai 200025, China; 2.Department of Imaging, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201999, China
  • Online:2016-11-28 Published:2016-11-29
  • Supported by:

    National Natural Science Foundation of China, 81602509; Shanghai Colleges and Universities Young Teachers Training Funding Scheme, ZZjdyx15016

摘要:

目的 ·探索线粒体二羧基载体蛋白SLC25A10对脑胶质瘤发生发展的作用及其机制。方法 ·通过慢病毒感染构建SLC25A10低表达和过表达的脑胶质瘤 U251细胞株。采用real-time PCR和Western blotting检测稳转U251细胞株中SLC25A10低表达和过表达的效率;通过细胞计数、CCK-8实验、克隆形成实验和裸鼠成瘤实验检测SLC25A10对脑胶质瘤细胞增殖和成瘤能力的影响;采用试剂盒和Western blotting分别检测细胞中的ATP含量和AMPK蛋白及其下游mTOR信号的活化情况。结果 · SLC25A10低表达后,U251细胞增殖减慢,克隆形成数目减少,成瘤能力和ATP含量亦降低。而SLC25A10过表达后,并未明显影响U251细胞的增殖,但增加了细胞ATP含量,并增强了细胞的放疗抵抗。机制研究表明SLC25A10低表达激活AMPK,抑制mTOR信号通路,从而降低细胞增殖能力,增强放疗敏感性。结论 · SLC25A10低表达抑制mTOR信号通路,减慢脑胶质瘤的恶性增殖,增强放疗敏感性。该发现可以为脑胶质瘤的临床治疗提供一个潜在的新靶点。

关键词: 脑胶质瘤, SLC25A10, 细胞增殖, ATP, mTOR

Abstract:

Objective · To explore the role and underlying mechanisms of mitochondrial dicarboxylate carrier SLC25A10 in the occurrence and progression of glioma. Methods · The U251 glioma cell lines over-expressing or depleted of SLC25A10 were obtained via lentivirus infection. Western blotting and real-time PCR were used to measure the efficiency of knockdown and over expression of SLC25A10. The effects of SLC25A10 on proliferation and tumorigenicity of U251 cells were detected with cell counting, CCK-8 assay, clonogenic assay, and tumor formation in nude mice. The ATP level and the activity of AMPK and its downstream mTOR signaling pathway were detected with kits and Western blotting, respectively. Results · Knock down of SLC25A10 decreased the proliferation, clone number, tumorigenicity, and ATP level in U251 cells. However, over expression of SLC25A10 didn’t significantly affect the proliferation of U251 cells, but increased the ATP level and the radiation resistance. Study showed that knock down of SLC25A10 activated AMPK, which in turn inhibited mTOR signaling pathway, resulting in decreased cell proliferation and increased cell sensitivity to radiotherapy. Conclusion · Knock down of SLC25A10 inhibits the mTOR signaling pathway, slows down glioma proliferation, and increases cell sensitivity to radiotherapy. These findings provide a potential target for the treatment of glioma.

Key words: glioma, SLC25A10, cell proliferation, ATP, mTOR