Analysis of Extracellular Vesicles in Gastric Juice from Gastric Cancer Patients
Analysis of Extracellular Vesicles in Gastric Juice from Gastric Cancer Patients
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DOI：
10.3390/ijms20040953
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发表时间：
2019-02-02
影响因子：
5.6
通讯作者：
Uchiyama, Kazuhisa
中科院分区：
文献类型：
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作者：
Kagota, Shuji;Taniguchi, Kohei;Uchiyama, Kazuhisa
细胞外小泡(EVS)是含有脂质、蛋白质和核酸的分泌膜小泡，它们通过向受体细胞运送它们的成分来发挥细胞间运输的功能。肠道病毒存在于各种体液中，如血液、唾液、尿液、羊水和腹水。癌细胞分泌的EV在其环境的形成中起着重要作用，包括纤维化、血管生成、逃避免疫监视，甚至转移。然而，胃液中的EVS(GJ-EVS)在很大程度上还没有被探索。在这项研究中，我们试图澄清来自胃癌患者的GJ-EVS的存在。用超速离心法结合我们自己的前处理方法从胃癌患者中分离出GJ-EVS。我们通过形态实验，即纳米粒子跟踪系统分析和电子显微镜对GJ-EVS进行了验证。此外，采用免疫印迹分析、生物分析仪或定量逆转录聚合酶链式反应检测EVS的蛋白质和microRNA标志物。GJ-EV能促进正常成纤维细胞的增殖。我们的发现表明，来自GC患者GJ的分离株含有EVS，这意味着GJ-EVS部分影响了他们的微环境，使用GC患者的GJ-EVS分析将有助于阐明GC的病理生理学。
Extracellular vesicles (EVs) are secretory membrane vesicles containing lipids, proteins, and nucleic acids; they function in intercellular transport by delivering their components to recipient cells. EVs are observed in various body fluids, i.e., blood, saliva, urine, amniotic fluid, and ascites. EVs secreted from cancer cells play important roles in the formation of their environment, including fibrosis, angiogenesis, evasion of immune surveillance, and even metastasis. However, EVs in gastric juice (GJ-EVs) have been largely unexplored. In this study, we sought to clarify the existence of GJ-EVs derived from gastric cancer patients. GJ-EVs were isolated by the ultracentrifuge method combined with our own preprocessing from gastric cancer (GC) patients. We verified GJ-EVs by morphological experiments, i.e., nanoparticle tracking system analysis and electron microscopy. In addition, protein and microRNA markers of EVs were examined by Western blotting analysis, Bioanalyzer, or quantitative reverse transcription polymerase chain reaction. GJ-EVs were found to promote the proliferation of normal fibroblast cells. Our findings suggest that isolates from the GJ of GC patients contain EVs and imply that GJ-EVs partially affect their microenvironments and that analysis using GJ-EVs from GC patients will help to clarify the pathophysiology of GC.