【摘要】 目的探究在人胶质瘤细胞系LN229中异柠檬酸脱氢酶1基因(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细胞株,分别对两组细胞株进行转录组测序(RNA-seq),利用limma包分析差异表达基因(DEGs)及其富集通路;然后通过string比对RBP数据库筛选出差异表达的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细胞株中,利用RNA-seq共筛选到259个DEGs,其中146个表达上调,113个表达下调;这些DEGs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的表达水平的改变无统计学意义。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。更多还原

【Abstract】 Objective To explore the role of IDH1 in the expression of RNA binding proteins( RBPs) in glioma cell line LN229. Methods Lentivirus was 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 was extracted for RNA sequencing( RNA-seq); the differentially expressed genes( DEGs) were identified by limma package 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 was tested in IDH1 knockdown,over-expressed and mutant cells( R132 H)respectively. Results A total of 259 DEGs were detected in IDH1 knockdown LN229 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 regulates expression of interferon pathway related gene IFIT3 in LN229 cells.更多还原