Screening of exosomal microRNAs from colorectal cancer cells
Glynn, Claire L.
Brown, James A.L.
Joyce, Myles R.
Kerin, Michael J.
Dwyer, Róisín M.
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Clancy, C,Khan, S,Glynn, CL,Holian, E,Dockery, P,Lalor, P,Brown, JAL,Joyce, MR,Kerin, MJ,Dwyer, RM (2016) 'Screening of exosomal microRNAs from colorectal cancer cells'. Cancer Biomarkers, 17 :427-435, DOI: 10.3233/CBM-160659
BACKGROUND: Cells release extracellular membrane vesicles including microvesicles known as exosomes. Exosomes contain microRNAs (miRNAs) however the full range within colorectal cancer cell secreted exosomes is unknown.OBJECTIVE: To identify the full range of exosome encapsulated miRNAs secreted from 2 colorectal cancer cell lines and to investigate engineering of exosomes over-expressing miRNAs.METHODS: Exosomes were isolated from HCT-116 and HT-29 cell lines. RNA was extracted from exosomes and microRNA array performed. Cells were engineered to express miR-379 (HCT-116-379) or a non-targeting control (HCT-116-NTC) and functional effects were determined. Exosomes secreted by engineered cells were transferred to recipient cells and the impact examined.RESULTS: Microvesicles 40-100 nm in size secreted by cell lines were visualised and confirmed to express exosomal protein CD63. HT-29 exosomes contained 409 miRNAs, HCT-116 exosomes contained 393, and 338 were common to exosomes from both cell lines. Selected targets were validated. HCT-116-379 cells showed decreased proliferation (12-15% decrease, p