Li Chunmei, Hou Xiaoming, Yuan Shuqiao, Zhang Yigan, Yuan Wenzhen, Liu Xiaoguang, Li Juan, Wang Yuping, Guan Quanlin, Zhou Yongning
Department of Gastroenterology, The First Hospital of Lanzhou University, Lanzhou, China.
Key Laboratory for Gastrointestinal Diseases of Gansu Province, The First Hospital of Lanzhou University, Lanzhou, China.
J Cancer. 2021 Apr 2;12(11):3277-3290. doi: 10.7150/jca.55077. eCollection 2021.
To date, the pathogenesis of gastric cancer (GC) remains unclear. We combined public database resources and bioinformatics analysis methods, explored some novel genes and verified the experiments to further understand the pathogenesis of GC and to provide a promising target for anti-tumor therapy. We downloaded the chip data related to GC from the Gene Expression Omnibus (GEO) database, extracted differentially expressed genes (DEGs), and then determined the key genes in the development of GC via PPI networks and model analysis. Functional annotation via GO and KEGG enrichment of DEGs was used to understand the latent roles of DEGs. The expression of the triggering receptor expressed on myeloid cells 2 (TREM2) gene in GC cell lines was verified via RT-PCR and western blotting. Moreover, the CCK-8, wound healing assay, and transwell migration and invasion assays were used to understand the changes in the proliferation, migration, and invasion abilities of GC cells after silencing TREM2. Western blotting verified the interaction between TREM2 and PI3K predict of the string website, as well as the effect of TREM2 on EMT. Finally, a lung metastasis model was used to explore the relationship between TREM2 and metastasis. Our study identified 16 key genes, namely BGN, COL1A1, COL4A1, COL5A2, NOX4, SPARC, HEYL, SPP1, TIMP1, CTHRC1, TREM2, SFRP4, FBXO32, GPX3, KIF4A, and MMP9 genes associated with GC. The EMT-related pathway was the most significantly altered pathway. TREM2 expression was higher in GC cell lines and was remarkably associated with tumor invasion depth, TNM stage, histological grade, histological type, anatomic subdivision, and state. Knockdown of TREM2 expression inhibited the proliferation, migration, and invasion of GC cells as well as the progression of EMT by PI3K/AKT signaling . In addition, lung metastasis were decreased . We identified some important genes associated with the progression of GC via public database analysis, explored and verified the effects of proto-oncogene TREM2 on EMT via the PI3K/AKT pathway. TREM2 may be a novel target in the GC therapy.
迄今为止，胃癌（GC）的发病机制仍不清楚。我们结合公共数据库资源和生物信息学分析方法，探索了一些新基因并进行实验验证，以进一步了解GC的发病机制，并为抗肿瘤治疗提供有前景的靶点。我们从基因表达综合数据库（GEO）下载了与GC相关的芯片数据，提取差异表达基因（DEGs），然后通过蛋白质-蛋白质相互作用（PPI）网络和模型分析确定GC发生发展中的关键基因。通过基因本体论（GO）和京都基因与基因组百科全书（KEGG）对DEGs进行功能注释，以了解DEGs的潜在作用。通过逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法验证髓系细胞触发受体2（TREM2）基因在GC细胞系中的表达。此外，采用细胞计数试剂盒-8（CCK-8）、伤口愈合实验以及Transwell迁移和侵袭实验，以了解沉默TREM2后GC细胞增殖、迁移和侵袭能力的变化。蛋白质免疫印迹法验证了TREM2与STRING网站预测的磷脂酰肌醇-3-激酶（PI3K）之间的相互作用，以及TREM2对上皮-间质转化（EMT）的影响。最后，利用肺转移模型探索TREM2与转移之间的关系。我们的研究确定了16个关键基因，即BGN、COL1A1、COL4A1、COL5A2、NOX4、SPARC、HEYL、SPP1、TIMP1、CTHRC1、TREM2、SFRP4、FBXO32、GPX3、KIF4A和MMP9基因与GC相关。EMT相关通路是改变最显著的通路。TREM2在GC细胞系中的表达较高，并且与肿瘤浸润深度、TNM分期、组织学分级、组织学类型、解剖亚部位和状态显著相关。敲低TREM2表达可抑制GC细胞的增殖、迁移和侵袭以及PI3K/AKT信号通路介导的EMT进程。此外，肺转移减少。我们通过公共数据库分析确定了一些与GC进展相关的重要基因，通过PI3K/AKT通路探索并验证了原癌基因TREM2对EMT的影响。TREM2可能是GC治疗中的一个新靶点。
