To Explore the Mechanism of the GRM4 Gene in Osteosarcoma by RNA Sequencing and Bioinformatics Approach.
To Explore the Mechanism of the GRM4 Gene in Osteosarcoma by RNA Sequencing and Bioinformatics Approach.
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利用RNA测序和生物信息学方法探讨GRM4基因在骨肉瘤中的作用机制
DOI：
10.12659/msmbr.908107
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发表时间：
2018-01-17
影响因子：
2.8
通讯作者：
He M
中科院分区：
文献类型：
--
作者：
Pang Y;Zhao J;Fowdur M;Liu Y;Wu H;He M
谷氨酸代谢受体4 （GRM4）与骨肉瘤的发病有关。本研究的目的是探讨GRM4在骨肉瘤中的潜在分子机制。材料/方法采用实时荧光定量PCR （RT-qPCR）检测4种人骨肉瘤细胞系和hFOB1.19细胞中GRM4的表达水平。用慢病毒介导的小干扰RNA （siRNA）转染GRM4表达最高的U2OS细胞。通过RNA测序筛选GRM4基因沉默后的差异表达基因（DEGs），并进行生物信息学分析。此外，利用生物信息学方法预测靶向GRM4的转录因子（tf），构建下游蛋白-蛋白相互作用（PPI）网络。结果共获得51个显著性基因，其中上调14个，下调37个。京都基因与基因组百科（KEGG）途径富集分析表明，获得了4条显著的富集途径。共检测到6个可能参与GRM4转录调控的tf。结果显示，PPI网络中的182个基因在14条通路上显著富集。趋化因子和趋化因子受体在三种途径中显著富集。结论四个显著富集通路中的deg可能通过GRM4参与骨肉瘤的发生发展。结果显示，EGR1和CTCF可能参与了GRM4的转录调控，GRM4通过与趋化因子及其受体的相互作用参与骨肉瘤的进展。
Background Glutamate metabotropic receptor 4 (GRM4) has been correlated with the pathogenesis of osteosarcoma. The objective of this study was to explore the underlying molecular mechanism of GRM4 in osteosarcoma. Material/Methods The expression levels of GRM4 in four human osteosarcoma cell lines and hFOB1.19 cells were examined by real-time quantitative PCR (RT-qPCR). The U2OS cells of the highest GRM4 expression were transfected with lentivirus-mediated small interfering RNA (siRNA). The differentially expressed genes (DEGs) after GRM4 gene silencing were screened through RNA sequencing, and analyzed by bioinformatics. Additionally, the transcription factors (TFs) targeting GRM4 were predicted and the downstream protein-protein interaction (PPI) network was constructed using the bioinformatics approach. Results A total of 51 significant DEGs were obtained, including 14 upregulated and 37 downregulated DEGs. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the DEGs indicated that four significant enrichment pathways were obtained. A total of six TFs that could be involved in the transcriptional regulation of GRM4 were detected. The results showed that 182 genes in the PPI network were significantly enriched in 14 pathways. The chemokines and chemokine receptors were found to be significantly enriched in three pathways. Conclusions The DEGs in the four significant enrichment pathways might participate in the development and progression of osteosarcoma through GRM4. The results revealed that EGR1 and CTCF are probably involved in the transcriptional regulation of GRM4, which participates in the progress of osteosarcoma by interacting with chemokines and their receptors.