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It has been reported that extracellular vesicles (EVs) are involved in the parasite-host interaction, and EVs of <jats:italic>C<\/jats:italic>. <jats:italic>sinensis<\/jats:italic> (<jats:italic>Cs<\/jats:italic>EVs) can contribute to biliary injuries and inflammation. However, uncertainty surrounds the function of <jats:italic>Cs<\/jats:italic>EVs in the progression of CCA. In this study, differential ultracentrifugation was used to separate <jats:italic>Cs<\/jats:italic>EVs from the culture supernatant of <jats:italic>C<\/jats:italic>. <jats:italic>sinensis<\/jats:italic> adult worms, and they were then identified by transmission electron microscopy, nanoparticle tracking analysis and proteome assays. CCK8, EdU-488 and colony formation assays were used to explore the effect of <jats:italic>Cs<\/jats:italic>EVs on the proliferation of CCA cells <jats:italic>in vitro<\/jats:italic>. Wound healing assays, transwell assays and <jats:italic>in vivo<\/jats:italic> lung metastasis model were conducted to evaluate the migration and invasion abilities. Moreover, the involvement of EMT process, as well as NF-\u03baB and ERK signaling pathway was assessed. Results showed that <jats:italic>Cs<\/jats:italic>EVs were successfully isolated and could be taken up by CCA cells, which promoted proliferation by accelerating cell cycle progression. In addition, <jats:italic>Cs<\/jats:italic>EVs could facilitate cell metastasis by triggering the epithelial-mesenchymal transition (EMT). Mechanistically, activation of NF-\u03baB signaling pathway was involved in the <jats:italic>Cs<\/jats:italic>EVs-mediated EMT, which could be reversed partly by BAY 11\u20137082 (an inhibitor of NF-\u03baB). In conclusion, these findings suggested that <jats:italic>Cs<\/jats:italic>EVs could induce the aberrant proliferation and metastasis of CCA cells by stimulating the NF-\u03baB\/EMT axis, providing a novel theoretical explanation for liver fluke-associated CCA.<\/jats:p>","DOI":"10.1371\/journal.pntd.0012545","type":"journal-article","created":{"date-parts":[[2024,10,28]],"date-time":"2024-10-28T17:40:56Z","timestamp":1730137256000},"page":"e0012545","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":10,"title":["Extracellular vesicles of Clonorchis sinensis promote the malignant phenotypes of cholangiocarcinoma via NF-\u03baB\/EMT axis"],"prefix":"10.1371","volume":"18","author":[{"given":"Xiaowen","family":"Pan","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qing","family":"He","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingxuan","family":"Yin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anyuan","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aoxun","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xueqing","family":"Yi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zifeng","family":"Zhong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yinjuan","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5518-1617","authenticated-orcid":true,"given":"Xuerong","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2024,10,28]]},"reference":[{"issue":"10020","key":"pntd.0012545.ref001","doi-asserted-by":"crossref","first-page":"800","DOI":"10.1016\/S0140-6736(15)60313-0","article-title":"Clonorchiasis","volume":"387","author":"MB Qian","year":"2016","journal-title":"Lancet"},{"issue":"1","key":"pntd.0012545.ref002","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s40249-016-0166-1","article-title":"Current status and perspectives of Clonorchis sinensis and clonorchiasis: epidemiology, pathogenesis, omics, prevention and control.","volume":"5","author":"ZL Tang","year":"2016","journal-title":"Infect Dis Poverty."},{"issue":"11","key":"pntd.0012545.ref003","doi-asserted-by":"crossref","first-page":"1014","DOI":"10.1016\/j.pt.2021.05.011","article-title":"Clonorchis sinensis.","volume":"37","author":"MB Qian","year":"2021","journal-title":"Trends Parasitol"},{"key":"pntd.0012545.ref004","doi-asserted-by":"crossref","first-page":"105309","DOI":"10.1016\/j.actatropica.2019.105309","article-title":"Clonorchis sinensis and clonorchiasis","volume":"203","author":"BK Na","year":"2020","journal-title":"Acta Trop"},{"issue":"3","key":"pntd.0012545.ref005","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.trecan.2015.08.011","article-title":"Why does infection with some helminths cause cancer?","volume":"1","author":"PJ Brindley","year":"2015","journal-title":"Trends Cancer."},{"issue":"1","key":"pntd.0012545.ref006","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1186\/s13045-022-01347-8","article-title":"The molecular mechanisms and therapeutic strategies of EMT in tumor progression and metastasis.","volume":"15","author":"Y Huang","year":"2022","journal-title":"J Hematol Oncol"},{"key":"pntd.0012545.ref007","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1146\/annurev-pathol-020117-043854","article-title":"Epithelial Mesenchymal Transition in Tumor Metastasis","volume":"13","author":"V. 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