基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 988-994.

• 研究论文 • 上一篇    下一篇

IDH1调控干扰素通路相关基因IFIT3的表达

唐婉军1,彭小忠2,韩为2   

  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 医学分子生物学国家重点实验室
  • 收稿日期:2021-04-20 修回日期:2021-05-18 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 韩为 E-mail:hanwei2012@ibms.pumc.edu.cn
  • 基金资助:
    国家自然科学基金

IDH1 regulates the expression of interferon pathway-related gene IFIT3

  • Received:2021-04-20 Revised:2021-05-18 Online:2021-07-05 Published:2021-06-17
  • Supported by:
    National Natural Science Foundation of China

摘要: 目的 探究在人胶质瘤细胞系LN229中IDH1对RNA结合蛋白(RBPs)表达的影响。方法 首先通过慢病毒感染构建IDH1稳定敲低和对照组LN229细胞系,分别对两组细胞系进行转录组测序(RNA-seq),分析差异表达基因及其富集通路;然后通过比对RBP数据库筛选出差异表达RBPs,构建蛋白相互作用网络并挑选出候选RBPs;最后在IDH1敲低、过表达和R132H突变的细胞系中进行表达量验证。结果 在IDH1敲低的LN229细胞系中,利用RNA-seq共筛选到259个差异表达基因,其中146个表达上调,113个表达下调;这些差异表达基因中包括14个编码RBP的基因,其中9个RBPs均为干扰素通路相关基因,并且具有密切的相互作用;与对照组相比,IDH1敲低时这9个RBPs表达水平均显著上调(P < 0.001), IDH1过表达时仅IFIT3表达显著下降(P < 0.01),而IDH1突变时IFIT3的表达水平未检测到明显改变。结论 在人胶质瘤细胞系LN229中,IDH1可调控干扰素通路相关基因IFIT3的表达。

关键词: 胶质瘤, IDH1, 干扰素通路, IFIT3

Abstract: Objective To explore the role of IDH1 in the expression of RNA binding proteins (RBPs) in glioma cell line LN229. Methods Lentiviruses were used to construct IDH1 knockdown and control cells, and the expression of IDH1 was validated by RT-qPCR (Real-time quantitative PCR) and Western blot. Then the total RNA of cells from each group were extracted for RNA sequencing (RNA-seq); further, the differentially expressed genes (DEGs) were identified by limma and enriched pathways were analyzed; the differentially expressed RBPs were selected according to the RBP database and protein-protein interaction network was constructed by String database; finally, the expression level of candidate RBPs were tested in IDH1 knockdown, overexpressed and mutant cells respectively. Results A total of 259 DEGs were detected in IDH1 knockdown cells, including 146 upregulated and 113 downregulated genes. These DEGs were significantly enriched in interferon related pathways and included 14 RBP-encoding genes, 9 of which were significantly enriched in interferon related pathways and showed close interaction; compared with the control group, IDH1 knockdown in LN229 significantly upregulated the expression of all the nine RBPs (P < 0.001), and IDH1 overexpression only downregulated IFIT3 (P < 0.01) involving in interferon pathway, while IDH1 mutation had no significant effect on IFIT3. Conclusions IDH1 could regulate the expression of interferon pathway related gene IFIT3 in glioma cell line LN229.

Key words: glioma, IDH1, interferon pathway, IFIT3