Li Tiankang, Zhou Tingting, Wu Jin, Lv Heng, Zhou Hui, Du Mingnan, Zhang Xiuzhong, Wu Nai, Gong Shuai, Ren Zeqiang, Zhang Pengbo, Zhang Chong, Liu Guangpu, Liu Xin, Zhang Yi
Department of General Surgery, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, China; Institute of Digestive Diseases, Xuzhou Medical University, Xuzhou, China.
Department of Endocrinology, The Affiliated Hospital of China University of Mining and Technology, Xuzhou, China; Department of Endocrinology, Xuzhou first People's Hospital, Xuzhou, China.
Transl Oncol. 2023 May;31:101652. doi: 10.1016/j.tranon.2023.101652. Epub 2023 Mar 17.
Although circular RNAs (circRNAs) have recently garnered interest as disease markers, they have been relatively poorly studied as a biomarker in colorectal cancer (CRC). In this study, we aimed to screen the exosome-derived circRNAs in CRC and explore their potential as diagnostic and prognostic biomarkers of CRC METHODS: Exosomes were extracted from the plasma using a kit and validated by immunoblotting, transmission electron microscopy, and particle size analysis. The microarray datasets were employed to identify differentially-expressed circRNAs from plasma exosomes. Real-time quantitative reverse transcription PCR (RT-qPCR) verified the results of the microarray analysis, and Receiver operating characteristic (ROC) curve revealed the diagnostic ability of a single circRNA. The Starbase combined with microT, miRmap, and RNA22 were used to establish a circRNA-miRNA-mRNA network. Gene ontology, Kyoto Encyclopedia of Genes, Genomes pathway enrichment analysis, and Gene Set Enrichment Analysis were applied to determine potential functions of the identified mRNAs RESULTS: Comparing the microarray of plasma exosome-derived circRNAs and the microarray downloaded from the GEO database, 15 candidate circRNAs with up-regulated expression were identified. RT-qPCR verified that hsa_circ_0003270 (circGAPVD1) was upregulated in CRC plasma exosomes. ROC analysis showed that circGAPVD1 in plasma exosomes has potential diagnostic value for CRC. The sensitivity and specificity of circGAPVD1 in the diagnosis of CRC were found to be 75.64 and 71.79%, respectively (area under ROC = 0.7662). Furthermore, the lymph node metastasis and TNM staging of patients were positively correlated with high expression of circGAPVD1. Combined with the ENCORI database and GEO datasets, we identified the circGAPVD1-related ceRNA network. The enrichment analysis revealed that key nodes in the ceRNA network participate in many important signaling pathways such as protein post-translational modifications CONCLUSION: Our results revealed the diagnostic efficiency of circGAPVD1 in plasma exosomes. The highly expressed circGAPVD1 is expected to be a novel diagnostic marker for CRC.
尽管环状RNA（circRNAs）最近作为疾病标志物引起了关注，但作为结直肠癌（CRC）的生物标志物，它们的研究相对较少。在本研究中，我们旨在筛选CRC中源自外泌体的circRNAs，并探索它们作为CRC诊断和预后生物标志物的潜力。方法：使用试剂盒从血浆中提取外泌体，并通过免疫印迹、透射电子显微镜和粒径分析进行验证。利用微阵列数据集鉴定血浆外泌体中差异表达的circRNAs。实时定量逆转录PCR（RT-qPCR）验证了微阵列分析的结果，受试者操作特征（ROC）曲线揭示了单个circRNA的诊断能力。使用Starbase结合microT、miRmap和RNA22建立circRNA-miRNA-mRNA网络。应用基因本体论、京都基因与基因组百科全书通路富集分析和基因集富集分析来确定已鉴定mRNA的潜在功能。结果：比较血浆外泌体来源的circRNAs微阵列和从GEO数据库下载的微阵列，鉴定出15个表达上调的候选circRNAs。RT-qPCR验证了hsa_circ_0003270（circGAPVD1）在CRC血浆外泌体中上调。ROC分析表明，血浆外泌体中的circGAPVD1对CRC具有潜在诊断价值。发现circGAPVD1在CRC诊断中的敏感性和特异性分别为75.64%和71.79%（ROC曲线下面积=0.7662）。此外，患者的淋巴结转移和TNM分期与circGAPVD1的高表达呈正相关。结合ENCORI数据库和GEO数据集，我们鉴定了circGAPVD1相关的ceRNA网络。富集分析表明，ceRNA网络中的关键节点参与许多重要信号通路，如蛋白质翻译后修饰。结论：我们的结果揭示了circGAPVD1在血浆外泌体中的诊断效率。高表达的circGAPVD1有望成为CRC的新型诊断标志物。
