earticle

영어

Exosomes (microvesicles, or micropaticles) are small membrane-enclosed vesicles and these are secreted by various cell types, including tumor cells. These vesicles play an important role as mediators in extracellular communication. They are composed of membrane, cellular proteins, DNA, and RNA derived from their origin cells. It is also known that exosomes are involved in tumor metastasis, angiogenesis, and antitumor immunity. These biological functions are probably due to the glycosylation on their membrane proteins. Thus, the study of glycosylation of exosomes will be another potential source of new biomarker. However, there is a little study about the glycosylation of exosomes. Here, we targeted and analyzed N-glycans of exosomes derived from five cancer cell lines (A549, PC9, PC9/ZD, MCF-7, and MDA-MB231) using nano-LC/MS. We also have compared glycans of exosomes with glycans on their origin cell membrane to examine glycosylation correlation between origin cells and exosomes on three lung cancer cell lines. Additionally, we have compared anticancer drug resistant cell line PC9/ZD (Gefitinib resistant) and untreated PC9 in terms of origin cells and exosomes glycan profiling via isomer separation. Exosomes and cell membranes were prepared from three lung cancer cell lines: A549, PC9, and PC9/ZD. And two breast cancer exosomes were prepared: MCF-7 and MDA-MB231. Glycans were released by PNGase F, then enriched by graphitized carbon solid phase extraction. Nano-LC/chip Q-TOF MS was used for overall glycan profiling and quantitation. We successfully release and profile N-glycans from exosomes. Origin cells and exosomes of lung cancer contain high-mannose glycans in abundance. Although, exosomes have less high-mannose glycans compared with origin cells. Both origin cells and exosomes, isomer separation of sialylated glycans are different between PC9 and PC9/ZD. On breast cancer exosomes, two different matastatic samples represent quite different profiling. This is the first study of comprehensive glycan profiling of exosomes using mass spectrometry.