Medical Laboratory, Taizhou Central Hospital (Taizhou University Hospital), Taizhou, Zhejiang, China.
Department of Medical Service, Taizhou Central Hospital (Taizhou University Hospital), Taizhou, Zhejiang, China.
Mediators Inflamm. 2021 Jan 7;2021:8819990. doi: 10.1155/2021/8819990. eCollection 2021.
Emerging evidence has shown that circular RNAs (circRNAs) and DNA methylation play important roles in the causation and progression of cancers. However, the roles of circRNAs and abnormal methylation genes in the tumorigenesis of pancreatic ductal adenocarcinoma (PDAC) are still largely unknown. Expression profiles of circRNA, gene methylation, and mRNA were downloaded from the GEO database, and differentially expressed genes were obtained via GEO2R, and a ceRNA network was constructed based on circRNA-miRNA pairs and miRNA-mRNA pairs. Inflammation-associated genes were collected from the GeneCards database. Then, functional enrichment analysis and protein-protein interaction (PPI) networks of inflammation-associated methylated expressed genes were investigated using Metascape and STRING databases, respectively, and visualized in Cytoscape. Hub genes of PPI networks were identified using the NetworkAnalyzer plugin. Also, we analyzed the methylation, protein expression levels, and prognostic value of hub genes in PDAC patients through the UALCAN, Human Protein Atlas (HPA), and Kaplan-Meier plotter databases, respectively. The circRNA_102049/miR-455-3p/CD80 axis was identified by the ceRNA network and hub genes. In vitro and in vivo experiments were performed to evaluate the functions of circRNA_102049. The regulatory mechanisms of circRNA_102049 and miR-455-3p were explored by RT-PCR, western blot, and dual-luciferase assays. In the present study, twelve hub genes (STAT1, CCND1, KRAS, CD80, ICAM1, ESR1, RAF1, RPS6KA2, KDM6B, TNRC6A, FOSB, and DNM1) were determined from the PPI networks. Additionally, the circRNA_102049 was upregulated in PDAC cell lines. Functionally, the knockdown of circRNA_102049 by siRNAs inhibited cell growth, inflammatory factors, and migratory and invasive potential and promoted cell apoptosis. Mechanistically, circRNA_102049 functioned as a sponge of miR-455-3p and partially reversed the effect of miR-455-3p and consequently upregulated CD80 expression. Our findings showed that circRNA_102049 and methylated hub genes play an important role in the proliferation, apoptosis, migration, invasion, and inflammatory response of PDAC, which might be selected as a promising prognostic marker and therapeutic target for PDAC.
越来越多的证据表明，环状 RNA（circRNA）和 DNA 甲基化在癌症的发生和发展中起着重要作用。然而，circRNA 和异常甲基化基因在胰腺导管腺癌（PDAC）的肿瘤发生中的作用仍知之甚少。从 GEO 数据库中下载 circRNA、基因甲基化和 mRNA 的表达谱，通过 GEO2R 获得差异表达基因，并基于 circRNA-miRNA 对和 miRNA-mRNA 对构建 ceRNA 网络。从 GeneCards 数据库中收集炎症相关基因。然后，分别使用 Metascape 和 STRING 数据库对炎症相关甲基化表达基因进行功能富集分析和蛋白质-蛋白质相互作用（PPI）网络分析，并在 Cytoscape 中可视化。使用 NetworkAnalyzer 插件识别 PPI 网络的枢纽基因。此外，我们分别通过 UALCAN、Human Protein Atlas（HPA）和 Kaplan-Meier plotter 数据库分析 PDAC 患者中枢纽基因的甲基化、蛋白表达水平和预后价值。通过 ceRNA 网络和枢纽基因鉴定 circRNA_102049/miR-455-3p/CD80 轴。通过体外和体内实验评估 circRNA_102049 的功能。通过 RT-PCR、western blot 和双荧光素酶报告基因实验探索 circRNA_102049 和 miR-455-3p 的调控机制。在本研究中，从 PPI 网络中确定了 12 个枢纽基因（STAT1、CCND1、KRAS、CD80、ICAM1、ESR1、RAF1、RPS6KA2、KDM6B、TNRC6A、FOSB 和 DNM1）。此外，circRNA_102049 在 PDAC 细胞系中上调。功能上，通过 siRNA 敲低 circRNA_102049 抑制细胞生长、炎症因子以及迁移和侵袭潜能，并促进细胞凋亡。机制上，circRNA_102049 作为 miR-455-3p 的海绵，部分逆转 miR-455-3p 的作用，进而上调 CD80 的表达。我们的研究结果表明，circRNA_102049 和甲基化枢纽基因在 PDAC 的增殖、凋亡、迁移、侵袭和炎症反应中发挥重要作用，可能作为 PDAC 有前途的预后标志物和治疗靶点。
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