细胞外囊泡（EVs）是由活细胞产生并分泌到细胞外基质中的双层膜囊泡。胆汁是肝细胞分泌的一种特殊体液，胆汁中是否存在外囊泡长链非编码RNA（exLRs）尚未得到探索。本研究对来自恶性肿瘤患者或胆道结石患者的19份胆汁样本进行了exLR测序（exLR-seq）分析。结果检测到了8649个mRNAs、13823个circRNAs和1105个lncRNAs。KEGG通路分析表明差异表达的exLRs富集在mTOR和AMPK信号通路中。我们鉴定出5个mRNAs（EID-2，LLPH，ATP6V0A2，RRP9和MTRNR2L10）、3个lncRNAs（AC015922.2，AL135905.1和LINC00921）和6个circRNAs（circASH1L，circATP9A，circCLIP1，circRNF138，circTIMMDC1和circANKRD12）富集在恶性肿瘤的胆汁EV样本中，这些exLRs可能是区分恶性肿瘤与良性胆道疾病的潜在标记物。此外，使用EV-origin算法对EV的组织/细胞来源成分进行了分析。结果显示，来自癌症患者的胆汁EV中CD4_naive和Th1细胞来源的绝对丰度显著增加。总之，我们的研究揭示了人体胆汁EV中丰富的exLRs，为选择肿瘤诊断标记物提供了一定依据。© 作者（们） 2023。由牛津大学出版社出版。保留所有权利。如需授权，请发送电子邮件至：journals.permissions@oup.com。

Extracellular vesicles (EVs) are bilayered membrane vesicles produced by living cells and secreted into the extracellular matrix. Bile is a special body fluid that is secreted by the liver cells, and extracellular vesicles long RNAs (exLRs) has not been explored in bile. In this study, exLR sequencing (exLR-seq) was performed on 19 bile samples from patients with malignant cancer or patients with biliary stones. A total of 8649 mRNAs, 13823 circRNAs and 1105 lncRNAs were detected. The KEGG pathway analysis revealed that differentially expressed exLRs were enriched in mTOR and AMPK signaling pathway. We identified five mRNAs (EID-2, LLPH, ATP6V0A2, RRP9, and MTRNR2L10), three lncRNAs (AC015922.2, AL135905.1, and LINC00921) and six circRNAs (circASH1L, circATP9A, circCLIP1, circRNF138, circTIMMDC1 and circANKRD12) were enriched in bile EV samples with cancer, and these exLRs may be potential markers used to distinguish malignant cancers from benign biliary diseases. Moreover, the tissue/cellular source components of EVs were analyzed using the EV-origin algorithm. The absolute abundance of CD4_naive and Th1 cell source in bile EVs from cancer patients were significantly increased. In summary, our study presented abundant exLRs in human bile EVs and provides some basis for the selection of tumor diagnostic markers.© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.