Yan Yulan, Zhang Riting, Zhang Xuanfeng, Zhang Anwei, Zhang Yao, Bu Xuefeng
Department of Respiratory Medicine, Affiliated People's Hospital of Jiangsu University Zhenjiang, Jiangsu, P. R. China.
Clinical Medicine College of Jiangsu University Zhenjiang, Jiangsu, P. R. China.
Am J Transl Res. 2019 Jan 15;11(1):160-175. eCollection 2019.
CircRNAs have been identified play a key role in various different types of cancer. However, their role in lung adenocarcinoma remains unclear. In this study, we explored the specific circular transcriptome and characterized the circRNA expression profiles of five paired lung adenocarcinoma (LAC) tissues relative to adjacent normal tissues from LAC patients using next-generation sequencing (NGS). To illuminate circRNAs function, their gene targets were initially predicted before using, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses to further analyse the associated significant cell signalling pathways and functions. The potential interactions between circRNA-miRNA-mRNA were also investigated. Additionally, qRT-PCR assay, Western blot and Immunohistochemistry were performed to validate the differential expression of circRNA, microRNA and mRNA in the LAC group in comparison to the control group. Two-hundred-eighty-five dysregulated circular transcripts were found in LAC tissues, among which 102 and 183 were either up or down regulated, respectively. Our biological analysis suggested that the host genes of differentially expressed circRNAs targeted to cancer-related processes and mechanisms. The interaction maps of the circRNA-miRNA-target gene were constructed using Cytoscope. In further study, hsa_circ_0002360 was found to be the most significantly overexpressed circRNA in LAC tissues by interacting with miRNA and its corresponding mRNA. Our results showed that hsa_circ_0002360 was aberrantly and abundantly expressed and implicated in the development of LAC, suggesting a valuable therapeutic target for LAC treatment.
环状RNA(circRNAs)已被证实参与多种不同类型癌症的发生发展过程。然而,它们在肺腺癌中的作用仍不明确。在本研究中,我们利用二代测序(NGS)技术对五对肺腺癌(LAC)患者的肿瘤组织及癌旁正常组织进行了环状转录组分析,以明确环状RNA的表达谱。为阐明环状RNA的功能,我们首先预测了其基因靶点,随后利用京都基因与基因组百科全书(KEGG)及基因本体论(GO)分析,进一步分析相关的重要细胞信号通路及功能。我们还研究了circRNA-miRNA-mRNA之间的潜在相互作用。此外,采用qRT-PCR、蛋白质免疫印迹法及免疫组化法对LAC组与对照组中circRNA、微小RNA(miRNA)及信使核糖核酸(mRNA)的差异表达进行验证。结果显示,LAC组织中共有285条环状转录本表达失调,其中102条上调,183条下调。生物学分析表明,差异表达的环状RNA宿主基因参与癌症相关的生物学过程及机制。利用Cytoscope构建了circRNA-miRNA-靶基因相互作用图谱。在进一步研究中,我们发现hsa_circ_0002360是LAC组织中表达上调最为显著的环状RNA,其通过与miRNA及其相应mRNA相互作用发挥作用。本研究结果表明,hsa_circ_0002360在LAC组织中异常高表达,参与LAC的发生发展,有望成为LAC治疗的重要靶点。