Proteomic analysis of exosomes from nasopharyngeal carcinoma cell identifies intercellular transfer of angiogenic proteins

Abstract

Exosomes, a group of secreted extracellular nanovesicles containing genetic materials and signaling molecules, play a critical role in intercellular communication. During tumorigenesis, exosomes have been demonstrated to promote tumor angiogenesis and metastasis while their biological functions in nasopharyngeal carcinoma (NPC) are poorly understood. In this study, we focused on the role of NPC-derived exosomes on angiogenesis. Exosomes derived from the NPC C666-1 cells and immortalized nasopharyngeal epithelial cells (NP69 and NP460) were isolated using ultracentrifugation. The molecular profile and biophysical characteristics of exosomes were verified by Western blotting, sucrose density gradient and electron microscopy. We showed that the C666-1 exosomes (10 and 20 lg/ml) could significantly increase the tubulogenesis, migration and invasion of human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner. Subsequently, an iTRAQ-based quantitative proteomics was used to identify the differentially expressed proteins in C666-1 exosomes. Among the 640 identified proteins, 51 and 89 proteins were considered as up-and down-regulated ( 1.5-fold variations) in C666-1 exosomes compared to the normal counterparts, respectively. As expected, pro-angiogenic proteins including intercellular adhesion molecule-1 (ICAM-1) and CD44 variant isoform 5 (CD44v5) are among the up-regulated proteins, whereas angio-suppressive protein, thrombospondin-1 (TSP-1) was down-regulated in C666-1 exosomes. Further confocal microscopic study and Western blotting clearly demonstrated that the alteration of ICAM-1 and TSP-1 expressions in recipient HUVECs are due to internalization of exosomes. Taken together, these data strongly indicated the critical roles of identified angiogenic proteins in the involvement of exosomes-induced angiogenesis, which could potentially be developed as therapeutic targets in future.Nasopharyngeal Carcinoma (NPC) is a common malignant tumor arising from the epithelium of nasopharynx and has a high prevalence in Southeast Asia, particularly in the Guangdong and Guangxi provinces. 1 NPC has a high rate of metastasis; and >60% of NPC patients are presented with cervical lymph node infiltration and 5-8% are presented with distal metastasis. 2 This high incidence of locoregional metastasis often resulted in the treatment failure and high mortality. 3 Therefore, exploring the mechanisms underlying NPC metastasis is indispensable for better management of the disease.In order to achieve tumor propagation and progression, induction of angiogenesis is one of the obligatory steps. With

Fetuin‐A (alpha 2HS glycoprotein) modulates growth, motility, invasion, and senescence in high‐grade astrocytomas

“…The present data support the notion that binding of fetuin-A-activated exosomes to their putative receptors on the cell surface transmits adhesion, motility, invasion, and growth signals in tumor cells [26,27,38]. The rapid in vivo growth of control LN229 cells also suggest that the secreted bioactive exosomes modified the tumor microenvironment by promoting angiogenesis [39,40]. This ensured the recruitment of more fetuin-A (liver derived) and other growth factors that accelerated the growth further.…”

Exosomes: Fighting Cancer With Cancer

“…Furthermore, HUVECs protein expressions of ICAM-1 and TSP-1 were found to be altered after internalisation of C666-1 exosomes. Consequently, angiogenesis was stimulated through induced tube formation, migration and invasion [143].…”

EFEMP1 as a Potential Biomarker for Diagnosis and Prognosis of Osteosarcoma

“…Progression in OS Mice. Tumor groups (10,20,30, and 35 days) for OS mice were established corresponding to 10-, 20-, 30-, and 35-day OS patients. Serum samples were collected at different time points (10, 20, 30, and 35 days, respectively) after tumor development.…”

“…Mechanistically, how EFEMP1 is released into the peripheral blood remains unclear. EFEMP1 has been reported in the study of exosome of nasopharyngeal carcinoma [20]. Then, it is likely that EFEMP1 is secreted either from the extracellular exosomes or from dying tumor tissues.…”