Xu Bei-Bei, Zheng En-Dian, Sun Hao-Yue, Huang Yi, Zheng Liang, Lan Qiao-Li, Zhou Xiao-Lu, Geng Xiao-Ge, Wang Ya-Nan, Wang Xiu-Yan, Yu Ying-Cong
Department of Gastroenterology, The Third Affiliated Hospital of Shanghai University, Wenzhou People's Hospital, Wenzhou Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou 325000, Zhejiang, China; Soochow University, Suzhou 215000, Jiangsu, China; Department of Gastroenterology, Zhejiang Provincial People's Hospital, Hangzhou 310000, Zhejiang, China.
Department of Gastroenterology, The Third Affiliated Hospital of Shanghai University, Wenzhou People's Hospital, Wenzhou Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou 325000, Zhejiang, China.
Transpl Immunol. 2023 Apr;77:101793. doi: 10.1016/j.trim.2023.101793. Epub 2023 Feb 10.
Circular RNA (circRNA) has been proved to be an important regulator of gastric cancer (GC). However, the role and regulatory mechanism of circrna related competitive endogenous RNA (ceRNA) in GC have not been established.
CircRNA data and clinical data were obtained from the GEO and TCGA databases. The ceRNA networks were constructed and a function enrichment analysis was completed. Additionally, correlations between hub genes expression, immune cell infiltration, and clinical phenotypes were determined. The differentially expressed circRNAs and their downstream microRNAs (miRNAs) were validated by quantitative real-time polymerase chain reaction, and the hub genes were validated by western blot analysis. The migration and invasion ability of overexpressed hsa_circ_0002504 was determined by a transwell assay.
The ceRNA network contained 2 circRNAs, 3 miRNAs, and 55 messenger RNAs (mRNAs). 323 biological processes terms, 53 cellular components terms, 51 molecular functions terms, and 4 signaling pathways were revealed by the function enrichment analysis. The GSEA analysis revealed that the hub genes were positively correlated with the axon guidance and adhesion molecules pathways. The correlation analysis revealed that overexpressed EPHA4 and KCNA1 indicated poor tissue differentiation and were associated with clinically advanced stages of GC. The in vitro experiments showed that hsa_circ_0002504 was significantly down-regulated in GC cell lines. In addition, the overexpression of hsa_circ_0002504 led to a significant downregulation of hsa-miR-615-5p and hsa-miR-767-5p, as well as an upregulation of EPHA4, KCNA1, and NCAM1. Furthermore, it suppressed the migration and invasion ability of GC cells.
Hsa_circ_0002504 is a potential diagnostic biomarker for GC. High expression of EPHA4 and KCNA1 may indicate poor prognosis.
环状RNA（circRNA）已被证明是胃癌（GC）的重要调节因子。然而，circRNA相关的竞争性内源RNA（ceRNA）在GC中的作用和调控机制尚未明确。
从GEO和TCGA数据库获取circRNA数据和临床数据。构建ceRNA网络并完成功能富集分析。此外，确定枢纽基因表达、免疫细胞浸润与临床表型之间的相关性。通过定量实时聚合酶链反应验证差异表达的circRNA及其下游微小RNA（miRNA），并通过蛋白质免疫印迹分析验证枢纽基因。通过Transwell实验确定过表达的hsa_circ_0002504的迁移和侵袭能力。
ceRNA网络包含2个circRNA、3个miRNA和55个信使RNA（mRNA）。功能富集分析揭示了323个生物学过程术语、53个细胞成分术语、51个分子功能术语和4条信号通路。基因集富集分析（GSEA）显示枢纽基因与轴突导向和黏附分子通路呈正相关。相关性分析显示，过表达的EPHA4和KCNA1表明组织分化差，且与GC的临床晚期相关。体外实验表明，hsa_circ_0002504在GC细胞系中显著下调。此外，hsa_circ_0002504的过表达导致hsa-miR-615-5p和hsa-miR-767-5p显著下调，以及EPHA4、KCNA1和NCAM1上调。此外，它抑制了GC细胞的迁移和侵袭能力。
Hsa_circ_0002504是GC的潜在诊断生物标志物。EPHA4和KCNA1的高表达可能预示预后不良。
