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Title: The role of extracellular vesicles in the oral cancer microenviroment
Author: Ofield, Mark
ISNI:       0000 0004 7226 8022
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2018
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Background: The oral cancer microenvironment is an evolving landscape comprised of different cell types. As the disease progresses environmental conditions within this landscape change and its composition must evolve and adapt to this for the tumour to develop. Since the discovery of mRNA and miRNA in extracellular vesicles (EVs) much interest has focused on the contribution of EVs to cancer. Here the role of EVs in oral cancer progression is explored. Methods: EVs were purified from the conditioned media of oral squamous carcinoma cells (OSCC) by serial centrifugation and size exclusion chromatography before characterisation with electron microscopy, western blotting and tuneable resistive pulse sensing. Next generation sequencing and mass spectrometry were used to identify the RNA and protein contents of these EVs. Bioinformatic approaches were used to correlate the contents with tumour stage of the producing cell. A fluorescence staining technique was used to image the transfer of EVs to cells of the tumour microenvironment in vitro and the impact of this uptake assessed in normal oral fibroblasts (NOF). Results: EVs of between 50-200 nm were produced by cells throughout oral cancer progression. These EVs were positive for markers CD63 and TSP1 but negative for the golgi body marker GM130. Transcriptomic and proteomic techniques have identified miRNA families and proteins changing in abundance in EVs with tumour stage including miR-200, miR-34, nespirin 1 and syndecan 4. EVs were taken up by cells of the tumour microenvironment. This uptake caused no change to NOF proliferation but activated them into a cancer associated fibroblast phenotype. Conclusions: EVs from OSCC are produced throughout tumorigenesis and their contents could reflect the status of the producing cell enabling them to act as diagnostic markers. EV uptake by NOFs caused them to become activated into a pro tumorigenic phenotype.
Supervisor: Lambert, Daniel ; Hunt, Stuart Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available