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血浆外泌体中早期肺腺癌环状 RNA 表达谱及潜在标志物

Circular RNAs expression profiles in plasma exosomes from early-stage lung adenocarcinoma and the potential biomarkers.

机构信息

Department of Clinical Laboratory, Fujian Provincial Hospital, Fuzhou, China.

Department of Clinical Laboratory, Shengli Clinical Medical College of Fujian Medical University, Fuzhou, China.

出版信息

J Cell Biochem. 2020 Mar;121(3):2525-2533. doi: 10.1002/jcb.29475. Epub 2019 Oct 23.

Abstract

This study aimed to identify differential circular RNA (circRNA) in the plasma exosomes of patients with lung adenocarcinoma (LUAD) using high-throughput sequencing. First, exosomes were isolated using an exosome isolation kit and confirmed by Western blotting, transmission electron microscopy, and NanoSight Assay. Subsequently, plasma circRNA expression profiles were screened by high-throughput sequencing and confirmed by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR) and Sanger sequencing. Finally, the circRNA-miRNA-mRNA network was performed to forecast the potential function of circRNAs. The result of high-throughput sequencing data documented that 182 differentially expressed exosomal circRNAs in all were screened, which included 105 that were upregulated and 78 that were downregulated in LUAD patients plasma compared with controls. The four upregulated circRNAs including circ_0001492, circ_0001346, circ_0000690, and circ_0001439 were identical to the sequencing data by qRT-PCR, and their latent circRNA-miRNA-mRNA interactions were exhibited. Taken together, our study firstly revealed the altered exosomal circRNA expression from plasma samples in patients with LUAD and supports the need for exploring their potential as biomarkers and the pathological effects of lung cancer.

摘要

本研究旨在通过高通量测序鉴定肺腺癌(LUAD)患者血浆外泌体中的差异环状 RNA(circRNA)。首先,使用外泌体分离试剂盒分离外泌体,并通过 Western blot、透射电子显微镜和 NanoSight 分析进行验证。随后,通过高通量测序筛选血浆 circRNA 表达谱,并通过荧光定量实时聚合酶链反应(qRT-PCR)和 Sanger 测序进行验证。最后,进行 circRNA-miRNA-mRNA 网络分析以预测 circRNAs 的潜在功能。高通量测序数据结果表明,共筛选出 182 个差异表达的外泌体 circRNA,其中 105 个在 LUAD 患者血浆中上调,78 个下调。qRT-PCR 验证了四个上调的 circRNA(circ_0001492、circ_0001346、circ_0000690 和 circ_0001439)与测序数据一致,并展示了它们潜在的 circRNA-miRNA-mRNA 相互作用。总之,本研究首次揭示了 LUAD 患者血浆样本中外泌体 circRNA 的改变表达,并支持探索其作为生物标志物的潜力以及肺癌的病理影响。

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