NDLI: Differential protein profiling of renal cell carcinoma urinary exosomes

Content Provider	Royal Society of Chemistry (RSC)
Author	Magni, F. Battaglia, C. Corbetta, S. Chinello, C. Bosari, S. Mina, P. Della Brambilla, P. Pitto, M. Albo, G. Morosi, L. Raimondo, F. Villa, A.
Copyright Year	2013
Abstract	Renal cell carcinoma (RCC) accounts for about 3% of all human malignancies and its incidence is increasing. There are no standard biomarkers currently used in the clinical management of patients with renal cell carcinoma. A promising strategy for new biomarker detection is comparative proteomics of urinary exosomes (UE), nanovesicles released by every epithelial cell facing the urinary space, enriched in renal proteins and excluding high-abundance plasmatic proteins, such as albumin. Aim of the work is to establish the protein profile of exosomes isolated from urines of RCC patient compared with control subjects. We enrolled 29 clear cell RCC patients and 23 control healthy subjects (CTRL), age and sex-matched, for urine collection and vesicle isolation by differential centrifugation. Such vesicles were morphologically and biochemically characterized and proved to share exosome properties. Proteomic analysis, performed on 9 urinary exosome (UE) pooled samples by gel based digestion followed by LC-MS/MS, led to the identification of 261 proteins from CTRL subject UE and 186 from RCC patient UE, and demonstrated that most of the identified proteins are membrane associated or cytoplasmic. Moreover, about a half of identified proteins are not shared between RCC and control UE. Starting from these observations, and from the literature, we selected a panel of 10 proteins, whose UE differential content was subjected to immunoblotting validation. Results show for the first time that RCC UE protein content is substantially and reproducibly different from control UE, and that these differences may provide clues for new RCC biomarker discovery.
Starting Page	1220
Ending Page	1233
Page Count	14
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	9
Issue Number	6
Journal	Molecular BioSystems
DOI	10.1039/c3mb25582d
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Urine Western blot Biomarker Carcinoma Albumin Epithelium Protein Exosome complex RCC CTRL Renal cell carcinoma Proteomics Differential centrifugation
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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