miR-26b-5p通过下调MKNK2/eIF4E信号通路抑制胰腺癌细胞的增殖及侵袭

MiR-26 b-5 p Downregulates Proliferation and Invasion of PANC-1 Cell via MKNK 2/eIF 4 E Signal Pathway

目的:筛选胰腺癌(PC)侵袭相关微小RNA(miRNA,miR),验证其生物学功能和调控的信号通路,为胰腺癌的诊治提供新的分子标志物和靶点。方法:通过基因表达综合数据库(GEO)的miRNA表达谱数据集GSE71533和GSE85589,分析miR-26b-5p的差异表达;利用癌症基因组图谱数据库(TCGA),分析miR-26b-5p对PC患者总生存率的影响。采用TargetScan数据库识别miR-26b-5p的靶基因集。通过细胞功能实验,验证miR-26b-5p对PANC-1细胞增殖、迁移和侵袭的影响。利用基因本体(GO)和KEGG通路富集分析,揭示miR-26b-5p靶基因的功能,并通过蛋白印迹(Western blotting)实验验证miR-26b-5p靶向的信号通路。结果:在GSE71533和GSE85589数据集,相对于正常组织和对照组血浆,miR-26b-5p在PC组织和PC患者血浆明显低表达(9.65±0.10 vs 10.18±0.07,P<0.001;0.67±0.18 vs 0.79±0.24,P=0.017)。TCGA-胰腺癌生存分析结果显示,miR-26b-5p高表达组的总体生存率明显优于低表达组(P=0.023)。KEGG分析显示,预测的251个miR-26b-5p的靶基因与MAPK信号通路有关。CCK-8实验、Transwell迁移、侵袭实验和划痕实验结果显示:miR-26b-5p抑制PANC-1细胞的增殖、迁移和侵袭。KEGG通路富集分析结果提示miR-26b-5p的靶基因参与丝裂原活化蛋白激酶(MAPK)信号通路。Western blotting实验证实miR-26b-5p下调MKNK2/eIF4E信号通路。结论:miR-26b-5p下调MKNK2/eIF4E信号通路,抑制PANC-1细胞增殖、侵袭,有可能作为胰腺癌诊断和治疗的新靶点。

Objective To screen out the microRNA associated with pancreatic cancer(PC)progression and validate its biological functions,target genes and potential signal pathways.Methods We performed differential analysis for miR-26b-5p,based on the GSE71533 and the GSE85589 datasets from Gene Expression Omnibus.Then,Kaplan-Meier survival analysis was conducted to evaluate the influence of miR-26b-5p on PC patients'overall survival based on the PC cohort of The Cancer Genome Atlas(TCGA).The TargetScan database was used to predict the target genes of miR-26b-5p and the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted to reveal the functions of the miR-26b-5p targeting gene set.in vitro and Western blotting assays were performed to validate the influence of miR-26b-5p on PANC-1 cell progression and to identify the target gene of miR-26b-5p.Results In GSE71533,miR-26b-5p was downregulated in PC tissues compared with normal tissues(9.65±0.10 vs 10.18±0.07,P<0.001).In GSE85589,miR-26b-5p was downregulated in PC patients'serum(0.67±0.18 vs 0.79±0.24,P=0.017).Downregulation of miR-26b-5p was associated with the poor overall survival in the PC cohort of TCGA(P=0.023).We predicted 251 target genes of miR-26b-5p through the Targetscan database and eight genes were involved in the MAPK signal pathway according to KEGG analysis.in vitro assays showed that miR-26b-5p suppressed PANC-1 cell proliferation,migration and invasion.Western blotting assays suggested that miR-26b-5p downregulated MKNK2 and the downstream gene eIF4E.Conclusion The miR-26b-5p suppressed PANC-1 cell proliferation and invasion via MKNK2/eIF4E.The miR-26b-5p could serve as a potential biomarker for PC diagnosis and a target for PC treatment.