Clonorchis sinensis-derived extracellular vesicle (CsEV) miRNA Csi-miR-190-3p exerts dual roles in cholangiocarcinoma progression via cell cycle modulation and CXCL5 signaling suppression

Abstract Clonorchis sinensis , a Class 1 carcinogen classified by International Agency for Research on Cancer (IARC), is strongly linked to cholangiocarcinoma (CCA) development. Extracellular vesicles of C. sinensis ( Cs EVs) facilitate parasite-host interactions by transferring microRNAs (miRNAs). This study investigated the role of Csi -miR-190-3p, a Cs EV-enriched miRNA, in regulating CCA progression. We transfected Csi -miR-190-3p mimics or inhibitors into RBE cells, and evaluated cellular functions (proliferation, migration, invasion) using CCK-8, EdU-488, colony formation, flow cytometry, wound healing and Transwell assays.A BALB/c nude mouse lung metastasis model was established to evaluate in vivo effects. Target genes were predicted using bioinformatics tools and validated by dual-luciferase reporter gene assays. RT-qPCR was employed to assess gene expression and signaling pathways. Results showed that Csi -miR-190-3p significantly promoted CCA cell proliferation via cell cycle acceleration, while potently inhibiting cell migration and invasion in vitro and in vivo. Csi -miR-190-3p directly bound to the 3′UTR of CXCL5 and downregulated its expression, thereby inhibiting metastasis. These findings uncover a novel dual regulatory mechanism of Cs EV-derived Csi -miR-190-3p in CCA progression, and identify Csi -miR-190-3p/CXCL5 axis as a potential therapeutic target for CCA.