Isolation and Characterization of Extracellular Vesicles from Gastric Juice

胃液中细胞外囊泡的分离和鉴定

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作者:Gleb O Skryabin, Svetlana V Vinokurova, Sergey A Galetsky, Danila S Elkin, Alexey M Senkovenko, Darya A Denisova, Andrey V Komelkov, Ivan S Stilidi, Ivan N Peregorodiev, Olga A Malikhova, Oiatiddin T Imaraliev, Adel D Enikeev, Elena M Tchevkina

Abstract

EVs are involved in local and distant intercellular communication and play a vital role in cancer development. Since EVs have been found in almost all body fluids, there are currently active attempts for their application in liquid diagnostics. Blood is the most commonly used source of EVs for the screening of cancer markers, although the percentage of tumor-derived EVs in the blood is extremely low. In contrast, GJ, as a local biofluid, is expected to be enriched with GC-associated EVs. However, EVs from GJ have never been applied for the screening and are underinvestigated overall. Here we show that EVs can be isolated from GJ by ultracentrifugation. TEM analysis showed high heterogeneity of GJ-derived EVs, including those with exosome-like size and morphology. In addition to morphological diversity, EVs from individual GJ samples differed in the composition of exosomal markers. We also show the presence of stomatin within GJ-derived EVs for the first time. The first conducted comparison of miRNA content in EVs from GC patients and healthy donors performed using a pilot sampling revealed the significant differences in several miRNAs (-135b-3p, -199a-3p, -451a). These results demonstrate the feasibility of the application of GJ-derived EVs for screening for miRNA GC markers.

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