OBJECTIVE: To observe the effect of exosomes released by adenoid cystic carcinoma (ACC) cell line SACC-83 on the proliferation of ACC cells. METHODS: Exosomes were isolated from SACC-83 cell culture supernatants using total exosome isolation reagents. The whole-mount exosomes were characterized using transmission electron microscope and Western blotting. The exosomes were labeled with green fluorescent dye PKH67 and co-cultured with SACC-83 cells for 48 h, followed by staining with Alexa Fluor 594 phalloidin and DAPI to observe exosome uptake by the cells using laser scanning confocal microscopy (LSCM). The cell proliferation was assessed using MTT assay and wound healing assay, and the expressions of ERK and P-ERK in the co-cultured SACC-83 cells were detected using Western blotting. RESULTS: The exosomes isolated from SACC-83 cells showed a size range of 30-100 nm and expressed the exosomal markers CD9, CD63 and TSG101. LSCM showed exosome uptake by SACC-83 cells, which exhibited accelerated proliferation and significantly enhanced P-ERK expression (P < 0.05) without significant changes in ERK expression. CONCLUSIONS: SACC-83 cells produce exosomes that promote the tumor cell proliferation and enhances the cellular expression of P-ERK, suggesting a potential role of MAPK/ERK pathway activation in exosome-mediated acceleration of ACC cell proliferation.
OBJECTIVE: To observe the effect of exosomes released by adenoid cystic carcinoma (ACC) cell line SACC-83 on the proliferation of ACC cells. METHODS: Exosomes were isolated from SACC-83 cell culture supernatants using total exosome isolation reagents. The whole-mount exosomes were characterized using transmission electron microscope and Western blotting. The exosomes were labeled with green fluorescent dye PKH67 and co-cultured with SACC-83 cells for 48 h, followed by staining with Alexa Fluor 594phalloidin and DAPI to observe exosome uptake by the cells using laser scanning confocal microscopy (LSCM). The cell proliferation was assessed using MTT assay and wound healing assay, and the expressions of ERK and P-ERK in the co-cultured SACC-83 cells were detected using Western blotting. RESULTS: The exosomes isolated from SACC-83 cells showed a size range of 30-100 nm and expressed the exosomal markers CD9, CD63 and TSG101. LSCM showed exosome uptake by SACC-83 cells, which exhibited accelerated proliferation and significantly enhanced P-ERK expression (P < 0.05) without significant changes in ERK expression. CONCLUSIONS: SACC-83 cells produce exosomes that promote the tumor cell proliferation and enhances the cellular expression of P-ERK, suggesting a potential role of MAPK/ERK pathway activation in exosome-mediated acceleration of ACC cell proliferation.
Authors: James A McCubrey; Linda S Steelman; William H Chappell; Stephen L Abrams; Ellis W T Wong; Fumin Chang; Brian Lehmann; David M Terrian; Michele Milella; Agostino Tafuri; Franca Stivala; Massimo Libra; Jorg Basecke; Camilla Evangelisti; Alberto M Martelli; Richard A Franklin Journal: Biochim Biophys Acta Date: 2006-10-07
Authors: R J Bryant; T Pawlowski; J W F Catto; G Marsden; R L Vessella; B Rhees; C Kuslich; T Visakorpi; F C Hamdy Journal: Br J Cancer Date: 2012-01-12 Impact factor: 7.640
Authors: Johan Skog; Tom Würdinger; Sjoerd van Rijn; Dimphna H Meijer; Laura Gainche; Miguel Sena-Esteves; William T Curry; Bob S Carter; Anna M Krichevsky; Xandra O Breakefield Journal: Nat Cell Biol Date: 2008-11-16 Impact factor: 28.824