Moldovan Leni, Batte Kara, Wang Yijie, Wisler Jon, Piper Melissa
Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Davis Heart and Lung Research Institute, Columbus, OH, USA.
Methods Mol Biol. 2013;1024:129-45. doi: 10.1007/978-1-62703-453-1_10.
Small extracellular vesicles are released from both healthy and disease cells to facilitate cellular communication. They have a wide variety of names including exosomes, microvesicles and microparticles. Depending on their size, very small extracellular vesicles originating from the endocytic pathway have been called exosomes and in some cases nanovesicles. Collectively, extracellular vesicles are important mediators of a wide variety of functions including immune cell development and homeostasis. Encapsulated in the extracellular vesicles are proteins and nucleic acids including mRNA and microRNA molecules. MicroRNAs are small, non-coding RNA molecules implicated in the post-transcriptional control of gene expression that have emerged as important regulatory molecules and are involved in disease pathogenesis including cancer. In some diseases, not only does the quantity and the subpopulations of extracellular vesicles change in the peripheral blood but also microRNAs. Here, we described the analysis of peripheral blood extracellular vesicles by flow cytometry and the RNA extraction from extracellular vesicles isolated from the plasma or serum to profile microRNA expression.
小细胞外囊泡由健康细胞和病变细胞释放，以促进细胞间通讯。它们有各种各样的名称，包括外泌体、微囊泡和微粒。根据其大小，源自内吞途径的非常小的细胞外囊泡被称为外泌体，在某些情况下也被称为纳米囊泡。总的来说，细胞外囊泡是多种功能的重要介质，包括免疫细胞发育和稳态。细胞外囊泡中包裹着蛋白质和核酸，包括mRNA和微小RNA分子。微小RNA是小的非编码RNA分子，参与基因表达的转录后调控，已成为重要的调节分子，并参与包括癌症在内的疾病发病机制。在某些疾病中，不仅外周血中细胞外囊泡的数量和亚群会发生变化，微小RNA也会发生变化。在这里，我们描述了通过流式细胞术分析外周血细胞外囊泡，以及从血浆或血清中分离的细胞外囊泡中提取RNA以分析微小RNA表达情况。
