engleski

PROTEOMIC ANALYSIS OF PLEURAL EFFUSION AND ITS POTENTIAL IN CLINICAL DIAGNOSTIC OF CANINE PULMONARY ADENOCARCINOMA

Patients with pleural, pericardial, and peritoneal effusions can be challenging, but effusion analysis can be an extremely useful tool as an inital step in making a diagnosis of malignancy or septic condition, both in human and veterinary medicine. There have been very few proteomic studies of effusions in veterinary medicine. Extracellular vesicles, like exosomes have an important role in intracellular communication, and may carry specific sets of proteins, lipids, miRNA, mRNA and DNA. In this study, we performed a semiquantitative characterization of exosomal marker proteins flotillin-1, adiponectin, VCAM- 1 and CD63 in two dogs with pulmonary adenocarcinoma. Samples of both blood serum and pleural effusion in studied patients and serum from control animal were analyzed. The exosomal proteins were tested by the Western blot method on the VWR Mini Vertical PAGE System (VWR International Ltd., Northern Ireland) and on the Electroblotting System (Mini Electroblotting System, Biovit, UK). Samples were loaded onto size exclusion chromatography columns (qEV, IZON Oxford) to separate extracellular vesicles from soluble proteins. Western blot of isolated extracellular vesicles showed significantly increased (p<0.05) serum levels of CD63, flotillin-1 and adiponectin in dogs with adenocarcinoma compared to healthy dogs. This finding could indicate that the prevalence of these extracellular vesicles is due to the tumor itself, or due to organisms reaction to the tumor itself. The presence of these vesicles in the pleural effusion could also be documented, but their levels were not indicative of the disease process. We found evidence that serum and effusions from patients with pulmonary adenocarcinoma contain extracellular vesicles with different protein composition. Further analysis of size comprehensive proteomic characterization in a larger population of patients could result in identification of potential biomarkers for the early detection of adenocarcinoma.

proteomic analysis, pleural effusion, adenocarcinoma

nije evidentirano