Identification of Serum Exosomal hsa-circ-0004771 as a Novel Diagnostic Biomarker of Colorectal Cancer
Identification of Serum Exosomal hsa-circ-0004771 as a Novel Diagnostic Biomarker of Colorectal Cancer
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DOI：
10.3389/fgene.2019.01096
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发表时间：
2019-11-01
影响因子：
3.7
通讯作者：
Wang, Shukui
中科院分区：
文献类型：
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作者：
Pan, Bei;Qin, Jian;Wang, Shukui
背景：外周血中的外泌体环状 RNA（circRNA）被认为是新兴的癌症诊断生物标志物。由于缺乏敏感和特异的生物标志物，大量结直肠癌（CRC）患者在诊断时已处于晚期，导致高死亡率。本研究旨在鉴定循环外泌体 circRNA 作为 CRC 的新型诊断生物标志物。材料和方法：通过将基因表达综合（GEO）数据库分析与在线程序 GEO2R 相结合来选择候选 circRNA。总共招募了 170 名患者和 45 名健康对照者来评估 circRNA 对 CRC 的诊断价值。通过透射电子显微镜（TEM）、纳米颗粒跟踪分析和蛋白质印迹证实从参与者血清和细胞培养基中分离出的外泌体。分别通过qRT-PCR和受试者工作特征（ROC）分析测试circRNA的表达和诊断效用。结果：根据GEO数据集分析结果，选择前10个差异表达circRNA中丰度最高（倍数变化>=1.5）的循环外泌体hsa-circ-0004771进行进一步研究。通过 qRT-PCR 验证，与健康对照 (HC) 和良性肠道疾病 (BID) 患者相比，CRC 患者血清中外泌体 hsa-circ-0004771 上调。循环外泌体 hsa-circ-0004771 的 ROC 曲线下面积 (AUC) 分别为 0.59 (95% CI, 0.457-0.725)、0.86 (95% CI, 0.785-0.933) 和 0.88 (95% CI, 0.815-0.940)，以区分 BID、I/II 期 CRC患者和结直肠癌患者 分别是 HC。 AUC 为 0.816（95%CI，0.728-0.9），可区分 I/II 期 CRC 患者与 BID 患者。此外，CRC患者血清中外泌体hsa-circ-0004771的表达升高是肿瘤源性的。结果发现，术后CRC患者血清以及经GW4869处理的CRC细胞培养基中外泌体hsa-circ-0004771的表达下调。结论：循环外泌体 hsa-circ-0004771 在 CRC 患者中显着上调，可作为 CRC 的新型潜在诊断生物标志物。
Background: Exosomal circular RNAs (circRNAs) in peripheral blood are considered as emerging diagnostic biomarkers of cancers. Owing to the lack of sensitive and specific biomarkers, a large number of colorectal cancer (CRC) patients were diagnosed in advanced stages leading to high mortality. This study aimed to identify circulating exosomal circRNAs as novel diagnostic biomarkers of CRC. Materials and Methods: Candidate circRNA was selected by integrating analysis of Gene Expression Omnibus (GEO) database with online program GEO2R. A total of 170 patients and 45 healthy controls were enrolled to assess the diagnostic value of circRNAs for CRC. Exosomes isolated from the serum of participants and cell cultured media were confirmed by transmission electron microscope (TEM), Nanoparticle Tracking Analysis and western blot. The expression and the diagnostic utility of circRNA were tested by qRT-PCR and receiver operating characteristic (ROC) analysis, respectively. Results: The circulating exosomal hsa-circ-0004771 with most abundant among the top ten differentially expressed circRNAs (fold change >= 1.5) was selected for further study based on the results of GEO dataset analysis. The up-regulated exosomal hsa-circ-0004771 was verified in serum of CRC patients compared to healthy controls (HCs) and patients with benign intestinal diseases (BIDs) by qRT-PCR. The area under the ROC curves (AUCs) of circulating exosomal hsa-circ-0004771 were 0.59 (95%CI, 0.457-0.725), 0.86 (95%CI, 0.785-0.933) and 0.88 (95%CI, 0.815-0.940) to differentiate BIDs, stage I/II CRC patients and CRC patients from HCs, respectively. The AUC was 0.816 (95%CI, 0.728-0.9) to differentiate stage I/II CRC patients from patients with BIDs. In addition, the elevated expression of exosomal hsa-circ-0004771 in the serum of CRC patients was tumor-derived. It was found that the expression of exosomal hsa-circ-0004771 was down-regulated expression of in the serum of postoperative CRC patients as well as cultured media of CRC cells treated with GW4869. Conclusions: Circulating exosomal hsa-circ-0004771 was significantly up-regulated in CRC patients and served as a novel potential diagnostic biomarker of CRC.