Interaction and uptake of exosomes by ovarian cancer cells

By Cristina Escrevente, Sascha Keller, P. Altevogt, Júlia Costa. BMC Cancer, 2011

629 citations recorded by the source corpus.

BackgroundExosomes consist of membrane vesicles that are secreted by several cell types, including tumors and have been found in biological fluids. Exosomes interact with other cells and may serve as vehicles for the transfer of protein and RNA among cells.MethodsSKOV3 exosomes were labelled with carboxyfluoresceine diacetate succinimidyl-ester and collected by ultracentrifugation. Uptake of these vesicles, under different conditions, by the same cells from where they originated was monitored by immunofluorescence microscopy and flow cytometry analysis. Lectin analysis was performed to investigate the glycosylation properties of proteins from exosomes and cellular extracts.ResultsIn this work, the ovarian carcinoma SKOV3 cell line has been shown to internalize exosomes from the same cells via several endocytic pathways that were strongly inhibited at 4°C, indicating their energy dependence. Partial colocalization with the endosome marker EEA1 and inhibition by chlorpromazine suggested the involvement of clathrin-dependent endocytosis. Furthermore, uptake inhibition in the presence of 5-ethyl-N-isopropyl amiloride, cytochalasin D and methyl-beta-cyclodextrin suggested the involvement of additional endocytic pathways. The uptake required proteins from the exosomes and from the cells since it was inhibited after proteinase K treatments. The exosomes were found to be enriched in specific mannose- and sialic acid-containing glycoproteins. Sialic acid removal caused a small but non-significant increase in uptake. Furthermore, the monosaccharides D-galactose, α-L-fucose, α-D-mannose, D-N-acetylglucosamine and the disaccharide β-lactose reduced exosomes uptake to a comparable extent as the control D-glucose.ConclusionsIn conclusion, exosomes are internalized by ovarian tumor cells via various endocytic pathways and proteins from exosomes and cells are required for uptake. On the other hand, exosomes are enriched in specific glycoproteins that may constitute exosome markers. This work contributes to the knowledge about the properties and dynamics of exosomes in cancer.

Subfields: Biotechnology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/9eeabc3cd597fd5bd62e3986e634b7e3d0af8b1b

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Citations in the corpus

Exosome Theranostics: Biology and Translational Medicine (2018, background): “Several studies have indicated that the tropism of exosomes might be determined by adhesion-associated molecules on the surface of exosomes, such as integrins, tetraspanins, and other glycoproteins [59].”
Biological properties of extracellular vesicles and their physiological functions (2015, background): “ferent between exosomes and apoptotic bodies (37).”
Exosomes and microvesicles: extracellular vesicles for genetic information transfer and gene therapy. (2012, background): “Some have hypothesized that exosomes and microvesicles can be phagocytosed by recipient cells (112,113) or fuse directly with the plasma membrane (114).”
Exosome and Exosomal MicroRNA: Trafficking, Sorting, and Function (2015, positive): “Escrevente et al. described a decrease in exosome uptake after the ovarian carcinoma cell line SKOV3 and its derived exosomes were treated with protease K, which indicated that the proteins mediating exosome internalization are presented on the surface of both the cells and the exosomes [40].”
Exosomes in cancer: small particle, big player (2015, background): “The uptake of exosomes by recipient cells is energy dependent [17].”
Routes and mechanisms of extracellular vesicle uptake (2014, background): “macropinocytosis (28,41), phagocytosis (22,27,41,82) and”
Overview of Extracellular Vesicles, Their Origin, Composition, Purpose, and Methods for Exosome Isolation and Analysis (2019, background): “Unlike exosomes and MVs, apoptotic bodies contain intact organelles, chromatin, and small amounts of glycosylated proteins [3,48,60,99].”