Dental Research Institute, University of California-Los Angeles, Los Angeles, CA 90095, USA.
Anal Chim Acta. 2012 Apr 20;723:61-7. doi: 10.1016/j.aca.2012.02.018. Epub 2012 Feb 19.
Microvesicles (MVs) have been shown to affect the physiology of neighboring recipient cells in various ways. They play an important role in tumor progression/metastasis and angiogenesis in cancer and may be useful therapeutic tools, as well as a mechanism of cell-to-cell communication. They have been visioned as an important biomarker or biomarker source for the detection of different diseases. Human saliva is a biological fluid with enormous diagnostic potential, which harbors plenty of salivary MVs. The goal of this study is to investigate the proteomic profiling of MVs in human saliva through a simple preparation procedure by using filtration and centrifugation. Gel electrophoresis was combined with LC-MS/MS (liquid chromatography-mass spectrometry) for the proteomic analysis of MVs. After SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) protein separation, the whole lane was cut into 25 bands, and each band was subjected to in-gel trypsin digestion. The peptides extracted from each band were loaded to LC-MS/MS for protein identification. Through protein database search, 63 proteins were identified for human salivary MVs. Several members of different protein families were identified, including annexin, keratin, actin, immunoglobulin and S100. This study showed that although there was an overlap with the proteins from human saliva and salivary exosomes, salivary MVs contained their own unique proteins. These results will poise human salivary MVs as a non-invasive tool for the early detection of different diseases.
微泡 (MVs) 已被证明以各种方式影响邻近受体细胞的生理功能。它们在癌症的肿瘤进展/转移和血管生成中发挥着重要作用，并且可能是有用的治疗工具，也是细胞间通讯的一种机制。它们被视为检测不同疾病的重要生物标志物或生物标志物来源。人唾液是一种具有巨大诊断潜力的生物液体，其中蕴藏着大量的唾液 MV。本研究的目的是通过使用过滤和离心的简单制备程序，研究人唾液中 MV 的蛋白质组特征。凝胶电泳与 LC-MS/MS（液相色谱-质谱）联合用于 MV 的蛋白质组分析。在 SDS-PAGE（十二烷基硫酸钠聚丙烯酰胺凝胶电泳）蛋白分离后，将整个泳道切成 25 个条带，每个条带进行胶内胰蛋白酶消化。从每个条带中提取的肽被加载到 LC-MS/MS 进行蛋白质鉴定。通过蛋白质数据库搜索，鉴定出 63 个人唾液 MV 的蛋白质。鉴定出了不同蛋白质家族的几个成员，包括膜联蛋白、角蛋白、肌动蛋白、免疫球蛋白和 S100。本研究表明，尽管人唾液和唾液外泌体中的蛋白质有重叠，但唾液 MV 含有其自身独特的蛋白质。这些结果将使人类唾液 MV 成为一种用于早期检测不同疾病的非侵入性工具。
