Identification and characterization of proteins isolated from microvesicles derived from human lung cancer pleural effusions

Abstract

Microvesicles (MVs, also known as exosomes, ectosomes, microparticles) are released by various cancer cells, including lung, colorectal, and prostate carcinoma cells. MVs released from tumor cells and other sources accumulate in the circulation and in pleural effusion. Although recent studies have shown that MVs play multiple roles in tumor progression, the potential pathological roles of MV in pleural effusion, and their protein composition, are still unknown. In this study, we report the first global proteomic analysis of highly purified MVs derived from human nonsmall cell lung cancer (NSCLC) pleural effusion. Using nano-LC-MS/MS following 1D SDS-PAGE separation, we identified a total of 912 MV proteins with high confidence. Three independent experiments on three patients showed that MV proteins from PE were distinct from MV obtained from other malignancies. Bioinformatics analyses of the MS data identified pathologically relevant proteins and potential diagnostic makers for NSCLC, including lung-enriched surface antigens and proteins related to epidermal growth factor receptor signaling. These findings provide new insight into the diverse functions of MVs in cancer progression and will aid in the development of novel diagnostic tools for NSCLC.

Proteomic study of benign and malignant pleural effusion

“…Proteins not previously reported were identified. Park et al (2013) reported the first proteomic analysis of purified microvesicles derived from In our study, we chose WCX-MB to purify peptides in the pleural effusion, used MALDI-TOF-MS to obtain peptide profiles from the pleural effusion samples, established and validated the diagnostic classification by GA CARD9 (Bertin et al 2000) was one of CARD-containing proteins, located on chromosome 9q34.3, and the cDNA with 2108 bp encoded a 536-amino acid protein. Human CARD9 was a novel protein with at least two functional domains.…”

Extracellular vesicles in lung microenvironment and pathogenesis

“…The global proteomic analysis of EVs derived from malignant pleural effusion of NSCLC showed that microvesicles contained 912 different proteins, including the epidermal growth factor receptor (EGFR), K-ras, basigin (EMMPRIN, CD147), carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), claudin1, claudin3, and RAB family proteins [78]. It has been reported that some EV proteins, such as leucine-rich a2-glycoprotein (LRG1), EGFR, CD91, and CD317, may be potential exosomal markers of NSCLC [79][80][81][82].…”

Characterization of Microvesicles Released from Human Red Blood Cells

“…However, these particles should be clearly separated based on other criteria rather than merely based on their size. However, so far efficient and specific markers to distinguish populations of MVs released from human RBCs have not been described [6,51].…”