上海交通大学学报(医学版) ›› 2017, Vol. 37 ›› Issue (6): 719-.doi: 10.3969/j.issn.1674-8115.2017.06.002

• 论著(基础研究) • 上一篇    下一篇

TRIM59 对神经胶质瘤形成的促进作用

宋丽娜,冯海忠   

  1. 上海交通大学 医学院附属仁济医院干细胞研究中心,上海 200127
  • 出版日期:2017-06-28 发布日期:2017-07-05
  • 通讯作者: 冯海忠,电子信箱:fenghaizhong@sjtu.edu.cn
  • 作者简介:宋丽娜(1990—),女,硕士生;电子信箱:songlina@sjtu.edu.cn
  • 基金资助:

    国家自然科学基金(81372704,81572467);上海高校特聘教授(东方学者)岗位计划(2014024);上海市教育委员会高峰高原学科建设计划(20161310)

Promotion of TRIM59 on neuroglioma genesis

SONG Li-na, FENG Hai-zhong   

  1. Clinical Stem Cell Research Center, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Online:2017-06-28 Published:2017-07-05
  • Supported by:

    National Natural Science Foundation of China, 81372704, 81572467; Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning, 2014024; Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support, 20161310

摘要:

] 目的 · 揭示TRIM59 在神经胶质瘤发生中的作用及分子机制。方法 · 通过免疫组织化学法分析TRIM59 蛋白在临床胶质瘤 手术切除样本中的表达;利用Western blotting 和定量PCR 法分析TRIM59 在多个胶质瘤细胞系中的表达;利用shRNA 敲低胶质瘤 细胞系中TRIM59 基因,分析其对胶质瘤细胞增殖、迁移、琼脂糖克隆形成及颅内原位肿瘤形成的影响;通过转录组测序及 KEGG PATHWAY 软件分析TRIM59 调节的信号通路。结果 · TRIM59 在神经胶质瘤组织中表达水平与肿瘤的临床分级呈正相关。相对于 正常星形胶质细胞,胶质瘤细胞系LN229 和 U87 细胞中TRIM59 表达较高。用慢病毒介导的shRNA 抑制U87 和 LN229 细胞中 TRIM59 的表达,能显著降低体外细胞增殖、迁移及克隆形成能力。用敲低 TRIM59 基因的 U87 细胞注射入裸鼠的右脑室,相较于对 照细胞,胶质瘤细胞体积明显缩小。转录组测序及KEGG PATHWAY 分析表明,敲低LN229 细胞TRIM59 基因后共有306 个基因下 调,其中与 PI3K/AKT 信号通路相关的基因最多。Western blotting 分析发现,LN229 和 U87 细胞敲低 TRIM59 后,AKT 磷酸化水平显 著降低,而过表达组成型活化的 Myr-AKT 可逆转 TRIM59 敲低对细胞增殖的抑制作用。结论 · TRIM59 是新发现的胶质瘤促癌基因, 其可能通过 PI3K/AKT 信号通路发挥作用。

关键词: &ensp, TRIM59, 胶质瘤, PI3K/AKT 信号通路, 促癌基因

Abstract:

 Objective · To demonstrate the effect of TRIM59 on gliomagenesis and the molecular mechanism.  Methods · TRIM59 protein expression in glioma specimens was analyzed by immunohistochemical staining, and in several glioma cell lines by Western blotting and quantative PCR. After TRIM59 was knocked down by shRNAs, cell proliferation, migration, colony formation, and orthotopic xenograft brain tumor development were detected. The signaling pathway of TRIM59 in gliomas was also explored by RNA-Seq and KEGG PATHWAY analyses.  Results · The levels of TRIM59 protein expression in clinical glioma specimens were positively correlated with glioma malignancy. TRIM59 was highly expressed in LN229 and U87 glioma cells compared with normal human astrocytes. Knockdown of TRIM59 in these two cell lines with lentivirus-mediated shRNAs inhibited their proliferation, migration, and colony formation. Compared with the control xenograft models, knockdown of TRIM59 significantly inhibited glioma tumor growth. RNASeq and KEGG PATHWAY analyses identified that TRIM59 knockdown down-regulated 306 genes, among which PI3K/AKT signal pathway-related genes were the most. Moreover, TRIM59 knockdown suppressed AKT phosphorylation, whereas overexpression of a constitutively actived AKT (Myr-AKT) rescued TRIM59 knockdown-inhibited cell proliferation.  Conclusion · TRIM59 is a new glioma oncogene, which may take effect through activating PI3K/ AKT signaling pathway.

Key words: TRIM59, glioma, PI3K/AKT signaling pathway, oncogene