Ruíz Martínez, MarcNavarro Ponz, AlfonsMarrades Sicart, Ramon Ma.Viñolas Segarra, NúriaSantasusagna, SandraMuñoz García, CarmenRamírez Ruz, J. (José)Molins López-Rodó, LaureanoMonzó Planella, Mariano2017-01-022017-01-022016-06-061949-2553https://hdl.handle.net/2445/105206BACKGROUND: Cancer-derived exosomes are involved in metastasis. YKT6 is a SNARE protein that participates in the regulation of exosome production and release, but its role in non-small cell lung cancer (NSCLC) has not been examined. MATERIALS AND METHODS: Ultracentrifugation-purified exosomes from the A549 cell line were studied by CRYO-TEM, nanoparticle tracking analysis and western blot (TSG101 marker). YKT6 was inhibited using a DsiRNA and selected pre-microRNAs. MicroRNAs targeting YKT6 were validated by Renilla/Luciferase assay and western blot. YKT6 expression and its prognostic impact were analyzed in 98 tissue specimens from resected NSCLC patients. RESULTS: Membranous nanosized vesicles (mode size: 128nm) with TSG101 protein were purified from A549 cells. YKT6 inhibition reduced exosome release by 80.9%. We validated miR-134 and miR-135b as miRNAs targeting YKT6, and transfection with the pre-miRNAs also produced a significant reduction in exosome release. The analysis of YKT6 in tumor samples showed that patients with high levels had shorter disease-free and overall survival. CONCLUSIONS: YKT6 is a key molecule in the regulation of exosome release in lung cancer cells and is in turn precisely regulated by miR-134 and miR-135b. Moreover, YKT6 levels impact prognosis of resected NSCLC patients.10 p.application/pdfengcc-by (c) Ruíz Martínez, Marc et al., 2016http://creativecommons.org/licenses/by/3.0/esCàncer de pulmóProteïnes citosquelètiquesMicro RNAsLung cancerCytoskeletal proteinsMicroRNAsinfo:eu-repo/semantics/article6632352017-01-02info:eu-repo/semantics/openAccess27285987