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• W2903300236 endingPage "140171" @default.
• W2903300236 startingPage "140171" @default.
• W2903300236 abstract "Extracellular vesicles comprise two main classes - exosomes and shed microvesicles (sMVs). Whilst much is known about exosome cargo content and functionality, sMVs are poorly understood. Here, we describe the large-scale purification of sMVs released from primary (SW480) and metastatic (SW620) human isogenic colorectal cancer (CRC) cell lines using a combination of differential ultracentrifugation and isopycnic iodixanol density centrifugation. The yield of SW480-sMVs and SW620-sMVs was 0.75 mg and 0.80 mg, respectively. Both SW480-/SW620-sMVs are heterogeneous in size (100-600 nm diameter) and exhibit identical buoyant densities (1.10 g/mL). In contrast to exosomes, sMVs are ALIX-, TSG101-, CD63- and CD9-. Quantitative mass spectrometry identified 1295 and 1300 proteins in SW480-sMVs and SW620-sMVs, respectively. Gene Ontology enrichment analysis identified 'cell adhesion' (CDH1, OCLN, CTN families), 'signalling pathway' (KRAS, NRAS, MAPK1, MAP2K1), and 'translation/RNA related' processes (EIF, RPL, HNRNP families) in both sMV types. Strikingly, SW480- and SW620-sMVs exhibit distinct protein signatures - SW480-sMVs being enriched in ITGA/B, ANXA1, CLDN7, CD44 and EGFR/NOTCH signalling networks, while SW620-sMVs are enriched in PRKCA, MACC1, FGFR4 and MTOR/MARCKS signalling networks. Both SW480- and SW620-sMVs are taken up by NIH3T3 fibroblasts resulting in similar cell invasion capability. This study provides, for the first time, molecular insights into sMVs and CRC biology." @default.
• W2903300236 created "2018-12-11" @default.
• W2903300236 creator A5005658403 @default.
• W2903300236 creator A5011373233 @default.
• W2903300236 creator A5013699003 @default.
• W2903300236 creator A5035048497 @default.
• W2903300236 creator A5082248830 @default.
• W2903300236 creator A5083100293 @default.
• W2903300236 date "2019-12-01" @default.
• W2903300236 modified "2023-10-10" @default.
• W2903300236 title "Proteomic profiling reveals key cancer progression modulators in shed microvesicles released from isogenic human primary and metastatic colorectal cancer cell lines" @default.
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