School of Clinical Medicine, St George and Sutherland Clinical Campuses, UNSW Sydney, Kensington, NSW 2052, Australia.
Cancer Care Centre, St George Hospital, Kogarah, NSW 2217, Australia.
Int J Mol Sci. 2023 Oct 23;24(20):15462. doi: 10.3390/ijms242015462.
Small extracellular vesicles (sEVs) are an important intercellular communicator, participating in all stages of cancer metastasis, immunity, and therapeutic resistance. Therefore, protein cargoes within sEVs are considered as a superior source for breast cancer (BC) biomarker discovery. Our study aimed to optimise the approach for sEV isolation and sEV proteomic analysis to identify potential sEV protein biomarkers for BC diagnosis. sEVs derived from BC cell lines, BC patients' plasma, and non-cancer controls were isolated using ultracentrifugation (UC), a Total Exosome Isolation kit (TEI), and a combined approach named UCT. In BC cell lines, the UC isolates showed a higher sEV purity and marker expression, as well as a higher number of sEV proteins. In BC plasma samples, the UCT isolates showed the highest proportion of sEV-related proteins and the lowest percentage of lipoprotein-related proteins. Our data suggest that the assessment of both the quantity and quality of sEV isolation methods is important in selecting the optimal approach for the specific sEV research purpose, depending on the sample types and downstream analysis.
小细胞外囊泡（sEVs）是一种重要的细胞间通讯者，参与癌症转移、免疫和治疗抵抗的所有阶段。因此，sEV 中的蛋白质货物被认为是乳腺癌（BC）生物标志物发现的优越来源。我们的研究旨在优化 sEV 分离方法和 sEV 蛋白质组学分析，以鉴定用于 BC 诊断的潜在 sEV 蛋白质生物标志物。使用超速离心（UC）、总外泌体分离试剂盒（TEI）和一种名为 UCT 的联合方法，从 BC 细胞系、BC 患者血浆和非癌症对照中分离 sEVs。在 BC 细胞系中，UC 分离物显示出更高的 sEV 纯度和标志物表达，以及更多的 sEV 蛋白。在 BC 血浆样本中，UCT 分离物显示出最高比例的 sEV 相关蛋白和最低比例的脂蛋白相关蛋白。我们的数据表明，评估 sEV 分离方法的数量和质量对于根据样品类型和下游分析选择最佳的特定 sEV 研究目的方法非常重要。
