Objective: To investigate the effect of exosomes derived from colon cancer (CC) cells and plasma of CC patients on migration of SW480 cells.
Methods: The exosomes derived from culture medium of human colon epithelial cell line NCM460 and CC cell line SW620 were isolated by ultracentrifugation. The exosomes derived from plasma of CC patients and healthy controls were isolated by size exclusion chromatography (SEC). The particle size and morphology of exosomes were identified by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) respectively, and exosomal markers were detected by Western blotting. The uptake of fluorescent DiI labeled exosomes by SW480 cells was observed by confocal microscopy. Transwell assay was used to detect the effect of exosomes on the migration of SW480 cells. The expression level of associated proteins in signaling pathway were analyzed by Western blotting. Rapamycin, an inhibitor of mTOR, was used to study the role of mTOR signaling pathway on exosomes mediated migration of SW480 cells.
Results: The results of NTA and TEM showed that the particle size of the isolated exosomes was about 120 nm, which were small vesicles with membrane structure. The expressions of exosomal markers Alix, TSG101 and CD63 could be detected. The exosomes were evidenced by a red fluorescent signal inside the cytoplasm of SW480 recipient cells, and could promote the migration of SW480 cells, which is associated with Akt/mTOR signaling pathway. Compared with the control group, plasma exosomes derived from CC patients could significantly promote the migration of SW480 cells. Inhibition the activity of mTOR signaling could attenuate the migration of SW480 cells.
Conclusions: Exosomes derived from CC cells and plasma of CC patients could promote the migration of SW480 cells, which is associated with Akt/mTOR signaling pathway.
Keywords: Akt/mTOR signaling pathway; Cell migration; Colon cancer; Exosomes.
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