Xin Wenxiu, Fu Yuxuan, Chen Liangsheng, Ding Haiying, Feng Tingting, Sun Jiao, Qi Yajun, Hu Yan, Fang Qilu, Fang Luo
Department of Pharmacy, Zhejiang Cancer Hospital, Hangzhou, China.
Wenzhou Medical University, Postgraduate Training Base of Zhejiang Cancer Hospital, Hangzhou, China.
J Gastrointest Oncol. 2023 Apr 29;14(2):719-732. doi: 10.21037/jgo-23-192.
In the progression of pancreatic ductal adenocarcinoma (PDAC), aberrant micro RNAs (miRNAs) expression plays a crucial role. This study sought to identify and validate the key miRNAs and potential target genes involved in PDAC. A bioinformatic analysis was conducted to determine their potential use as biomarkers and therapeutic targets.
Gene profiling data sets (GSE41372 and GSE32688) were retrieved from the Gene Expression Omnibus database. Differentially expressed miRNAs (DEMs) with a P value <0.05, and |fold change| >2 was identified. The prognostic value of the DEMs was accessed using the online server Kaplan-Meier plotter. Further, gene ontology terms and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed using DAVID 6.7. The protein-protein interaction analyses were conducted with STRING, and miRNA-hub gene networks were constructed using Cytoscape software. The PDAC cells were transfected with miRNA inhibitors or mimics. Cell Counting Kit-8 assays and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were used to examine cell proliferation and apoptosis, respectively. Wound-healing assays were performed to evaluate cell migration.
Three DEMs (hsa-miR-21-5p, hsa-miR-135b-5p, and hsa-miR-222-3p) were identified. High expression levels of hsa-miR-21-5p, hsa-miR-135b-5p, or hsa-miR-222-3p predicted poor overall survival in PDAC patients. The pathway analysis revealed that the predicted target genes of the DEMs were closely related to several signaling pathways (including 'pathways in cancer', 'miRNAs in cancer', 'platinum drug resistance', 'lipid and atherosclerosis', and 'MAPK signaling pathway'). The MYC proto-oncogene (), phosphate and tensin homolog gene (), poly(ADP-ribose) polymerase 1 (), von Hippel-Lindau (), and fork head box p3 () were identified as potential target genes. The inhibition of hsa-miR-21-5p, hsa-miR-135b-5p, or hsa-miR-222-3p expression decreased cell proliferation. The overexpression of hsa-miR-21-5p, hsa-miR-135b-5p, or hsa-miR-222-3p facilitated PDAC cell migration.
This study constructed the miRNA-hub gene network, which provides novel insights into the PDAC progression. Although further research is required, our results offer clues for new potential prognostic markers and therapeutic targets of PDAC.
在胰腺导管腺癌（PDAC）的进展过程中，异常的微小RNA（miRNA）表达起着关键作用。本研究旨在鉴定和验证参与PDAC的关键miRNA和潜在靶基因。进行了生物信息学分析以确定它们作为生物标志物和治疗靶点的潜在用途。
从基因表达综合数据库中检索基因谱数据集（GSE41372和GSE32688）。鉴定出P值<0.05且|倍数变化|>2的差异表达miRNA（DEM）。使用在线服务器Kaplan-Meier绘图仪评估DEM的预后价值。此外，使用DAVID 6.7进行基因本体术语和京都基因与基因组百科全书通路分析。使用STRING进行蛋白质-蛋白质相互作用分析，并使用Cytoscape软件构建miRNA-枢纽基因网络。用miRNA抑制剂或模拟物转染PDAC细胞。分别使用细胞计数试剂盒-8测定法和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色来检测细胞增殖和凋亡。进行伤口愈合试验以评估细胞迁移。
鉴定出3种DEM（hsa-miR-21-5p、hsa-miR-135b-5p和hsa-miR-222-3p）。hsa-miR-21-5p、hsa-miR-135b-5p或hsa-miR-222-3p的高表达水平预示着PDAC患者的总生存期较差。通路分析显示，DEM的预测靶基因与几种信号通路密切相关（包括“癌症中的通路”、“癌症中的miRNA”、“铂类耐药”、“脂质与动脉粥样硬化”和“MAPK信号通路”）。MYC原癌基因、磷酸酶和张力蛋白同源基因、聚（ADP-核糖）聚合酶1、冯·希佩尔-林道基因和叉头框p3被鉴定为潜在靶基因。抑制hsa-miR-21-5p、hsa-miR-135b-5p或hsa-miR-222-3p的表达可降低细胞增殖。hsa-miR-
