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长非编码 RNA LINC00968 在非小细胞肺癌 A549 细胞中的 lncRNA-miRNA-mRNA 调控网络的前瞻性研究:miRNA 微阵列和生物信息学研究。

Prospective lncRNA-miRNA-mRNA regulatory network of long non-coding RNA LINC00968 in non-small cell lung cancer A549 cells: A miRNA microarray and bioinformatics investigation.

机构信息

Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.

Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.

出版信息

Int J Mol Med. 2017 Dec;40(6):1895-1906. doi: 10.3892/ijmm.2017.3187. Epub 2017 Oct 12.

Abstract

Accumulating evidence suggests that the dysregulation of long non-coding RNAs (lncRNAs) serves vital roles in the incidence and progression of lung cancer. However, the molecular mechanisms of LINC00968, a recently identified lncRNA, remain unknown. The objective of present study was to investigate the role of a prospective lncRNA-miRNA‑mRNA network regulated by LINC00968 in non-small cell lung cancer cells. Following the transfection of lentiviruses carrying LINC00968 into A549 cells, the microRNA (miRNA) expression profile of the cells in response to the overexpression of LINC00968 was detected using an miRNA microarray. Five differentially expressed miRNAs (DEMs) with LINC00968 overexpression were obtained, including miR-9-3p, miR‑22-5p, miR-668-3p, miR‑3675-3p and miR-4536-3p. Five target prediction algorithms and three target validation algorithms were used to obtain 1,888 prospective target genes of the five DEMs. The result of Gene Ontology analysis suggested that these five DEMs were involved in complex cellular pathways, which included intracellular transport, organelle lumen and nucleotide binding. Furthermore, analysis of Kyoto Encyclopedia of Genes and Genomes pathways indicated that the five DEMs were important regulators in the adherens junction and focal adhesion. An lncRNA-miRNA-mRNA regulatory network and a protein-protein interaction network were then constructed. Eventually, a prospective lncRNA‑miRNA-mRNA regulatory network of LINC00968, three miRNAs (miR-9, miR-22 and miR-4536) and two genes (polo-like kinase 1 and exportin-1) was obtained following validation in the Cancer Genome Atlas database. These results may provide novel insights to support future research into lncRNA in lung cancer.

摘要

越来越多的证据表明,长非编码 RNA(lncRNA)的失调在肺癌的发生和发展中起着至关重要的作用。然而,最近发现的 lncRNA LINC00968 的分子机制尚不清楚。本研究旨在探讨由 LINC00968 调控的 lncRNA-miRNA-mRNA 网络在非小细胞肺癌细胞中的作用。用携带 LINC00968 的慢病毒转染 A549 细胞后,用 miRNA 微阵列检测细胞对 LINC00968 过表达的 miRNA 表达谱。获得了 5 个差异表达 miRNA(DEMs),包括 miR-9-3p、miR-22-5p、miR-668-3p、miR-3675-3p 和 miR-4536-3p。使用 5 种靶基因预测算法和 3 种靶基因验证算法,获得了这 5 个 DEMs 的 1888 个潜在靶基因。GO 分析结果表明,这 5 个 DEMs 参与了复杂的细胞通路,包括细胞内运输、细胞器腔和核苷酸结合。此外,KEGG 通路分析表明,这 5 个 DEMs 是黏着斑和焦点黏附的重要调节因子。然后构建了 lncRNA-miRNA-mRNA 调控网络和蛋白质-蛋白质相互作用网络。最终,在癌症基因组图谱数据库中进行验证后,获得了 LINC00968、3 个 miRNA(miR-9、miR-22 和 miR-4536)和 2 个基因(polo 样激酶 1 和 exportin-1)的 lncRNA-miRNA-mRNA 调控网络。这些结果可能为未来的肺癌 lncRNA 研究提供新的见解。

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