Department of Rehabilitation, Shanghai General Hospital, Shanghai Jiao Tong University, No. 100, Haining Road, Shanghai, 200080, China.
School of International Medical Technology, Shanghai Sanda University, No. 2727, Jinhai Road, Shanghai, 201209, China.
Biol Res. 2018 Aug 20;51(1):26. doi: 10.1186/s40659-018-0175-6.
Diffuse intrinsic pontine glioma (DIPG) is the main cause of pediatric brain tumor death. This study was designed to identify key genes associated with DIPG.
The gene expression profile GSE50021, which consisted of 35 pediatric DIPG samples and 10 normal brain samples, was downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) were identified by limma package. Functional and pathway enrichment analyses were performed by the DAVID tool. Protein-protein interaction (PPI) network, and transcription factor (TF)-microRNA (miRNA)-target gene network were constructed using Cytoscape. Moreover, the expression levels of several genes were validated in human glioma cell line U251 and normal glia HEB cells through real-time polymerase chain reaction (PCR).
A total of 378 DEGs were screened (74 up-regulated and 304 down-regulated genes). In the PPI network, GRM1, HTR2A, GRM7 and GRM2 had higher degrees. Besides, GRM1 and HTR2A were significantly enriched in the neuroactive ligand-receptor interaction pathway, and calcium signaling pathway. In addition, TFAP2C was a significant down-regulated functional gene and hsa-miR-26b-5p had a higher degree in the TF-miRNA-target gene network. PCR analysis revealed that GRM7 and HTR2A were significantly downregulated while TFAP2C was upregulated in U251 cells compared with that in HEB cells (p < 0.001). GRM2 was not detected in cells.
GRM1 and HTR2A might function in DIPG through the neuroactive ligand-receptor interaction pathway and the calcium signaling pathway. Furthermore, the TFAP2C and hsa-miR-26b-5p might play important roles in the development and progression mechanisms of DIPG.
弥漫性内在脑桥胶质瘤（DIPG）是小儿脑肿瘤死亡的主要原因。本研究旨在鉴定与 DIPG 相关的关键基因。
从基因表达综合数据库中下载包含 35 例小儿 DIPG 样本和 10 例正常脑组织样本的基因表达谱数据集 GSE50021。使用 limma 包鉴定差异表达基因（DEGs）。使用 DAVID 工具进行功能和通路富集分析。使用 Cytoscape 构建蛋白质-蛋白质相互作用（PPI）网络和转录因子（TF）-微小 RNA（miRNA）-靶基因网络。此外，通过实时聚合酶链反应（PCR）验证了几个基因在人神经胶质瘤细胞系 U251 和正常胶质细胞 HEB 中的表达水平。
筛选出 378 个差异表达基因（74 个上调和 304 个下调基因）。在 PPI 网络中，GRM1、HTR2A、GRM7 和 GRM2 的节点度较高。此外，GRM1 和 HTR2A 显著富集在神经活性配体-受体相互作用途径和钙信号通路中。此外，TFAP2C 是一个显著下调的功能基因，在 TF-miRNA-靶基因网络中 hsa-miR-26b-5p 的节点度较高。PCR 分析显示，与 HEB 细胞相比，U251 细胞中 GRM7 和 HTR2A 显著下调，而 TFAP2C 上调（p<0.001）。细胞中未检测到 GRM2。
GRM1 和 HTR2A 可能通过神经活性配体-受体相互作用途径和钙信号通路在 DIPG 中发挥作用。此外，TFAP2C 和 hsa-miR-26b-5p 可能在 DIPG 的发生和发展机制中发挥重要作用。
