Zhang Yalu, Zhang Rundong, Ding Xi, Ai Kaixing
Department of General Surgery, Tongji Hospital, School of Medicine, Tongji University Shanghai 200065, China.
Department of General Surgery, Shanghai Pulmonary Hospital, School of Medicine, Tongji University Shanghai 200433, China.
Am J Transl Res. 2018 Nov 15;10(11):3514-3528. eCollection 2018.
This study aims to identify the pivotal microRNAs (miRNAs) and genes, and their potential regulatory mechanisms in pancreatic ductal adenocarcinoma (PDAC) through bioinformatics analysis and experimental verification. We comprehensively analyzed two miRNA microarray datasets (GSE32678 and GSE43796) and three gene microarray datasets (GSE28735, GSE41368 and GSE71989), which were downloaded from the Gene Expression Omnibus (GEO) database, and identified the total of 8 differentially expressed miRNAs (DEMs) and 257 differentially expressed genes (DEGs) in common. Next, a new miRNA-mRNA regulatory network was constructed by bioinformatics methods, including 7 miRNAs, 58 putative target genes and 80 interaction pairs of miRNA-mRNA. Scrutinized by OncoLnc and GEPIA, it was found that 3 of 7 miRNAs (miR-21, miR-196b and miR-203) and 20 of 58 genes (MXRA5, EPYC, ECT2, COL12A1, SLC6A14, SLC7A2, BTG2, PDK4, CTNND2, NRP2, PXDN, CD109, TGFBI, LRRN1, ITGA2, DKK1, GREM1, EFNB2, SEMA3C and NT5E) were notably associated with prognosis in patients with PDAC. Furthermore, EFNB2 was significantly upregulated in PDAC compared with normal controls from different public databases. Cellular function experiments demonstrated that EFNB2 knockdown inhibited cell proliferation, migration and invasion in SW1990 cells. Western blot and luciferase reporter assays revealed that miR-557 negatively regulated the expression of EFNB2 by directly binging its 3' UTR. In conclusion, we performed integrated analysis for multiple expression profiles, and provided novel candidate miRNAs and genes to be exploited for functional studies. In addition, our findings suggested that EFNB2 contributes to PDAC progression by acting as the target gene of miR-557. It is useful for uncovering miRNA-based treatments in PDAC.
本研究旨在通过生物信息学分析和实验验证,确定胰腺导管腺癌(PDAC)中的关键微小RNA(miRNA)和基因及其潜在调控机制。我们全面分析了从基因表达综合数据库(GEO)下载的两个miRNA微阵列数据集(GSE32678和GSE43796)以及三个基因微阵列数据集(GSE28735、GSE41368和GSE71989),共鉴定出8个差异表达的miRNA(DEM)和257个差异表达基因(DEG)。接下来,通过生物信息学方法构建了一个新的miRNA - mRNA调控网络,包括7个miRNA、58个假定的靶基因和80个miRNA - mRNA相互作用对。经OncoLnc和GEPIA分析发现,7个miRNA中的3个(miR - 21、miR - 196b和miR - 203)以及58个基因中的20个(MXRA5、EPYC、ECT2、COL12A1、SLC6A14、SLC7A2、BTG2、PDK4、CTNND2、NRP2、PXDN、CD109、TGFBI、LRRN1、ITGA2、DKK1、GREM1、EFNB2、SEMA3C和NT5E)与PDAC患者的预后显著相关。此外,从不同公共数据库可知,与正常对照相比,EFNB2在PDAC中显著上调。细胞功能实验表明,敲低EFNB2可抑制SW1990细胞的增殖、迁移和侵袭。蛋白质免疫印迹和荧光素酶报告基因检测显示,miR - 557通过直接结合EFNB2的3'非翻译区负向调控其表达。总之,我们对多个表达谱进行了综合分析,为功能研究提供了新的候选miRNA和基因。此外,我们的研究结果表明,EFNB2作为miR - 557的靶基因促进了PDAC的进展。这对于揭示基于miRNA的PDAC治疗方法具有重要意义。