尿细胞外囊泡（uEV）反映了产生细胞的生物学状况。 uEV 的蛋白质分析使我们能够更好地了解膀胱癌、前列腺癌和肾癌等几种癌症的癌症进展，但在乳腺癌中尚未有报道。在此，我们的目的是量化乳腺癌 (BC) 患者与健康对照 (CT) 患者中 uEV 的浓度并生成蛋白质组谱。尿样采集自 29 名 CT 患者和 47 名 BC 患者。使用差速超速离心分离 uEV，然后通过蛋白质印迹和透射电子显微镜进行表征。此外，还使用纳米颗粒跟踪分析来测量尿液颗粒和 uEV 的浓度和尺寸分布。通过 LC-MS/MS 促进了 uEV 的蛋白质组分析。评估组之间的 uEV 浓度没有显着差异。进行的蛋白质组分析显示 BC 患者组和 CT 组分别有 15,473 和 11,278 个蛋白质。此外，热图分析揭示了差异蛋白表达，而主成分分析则突出显示了两个簇。火山图表明，与 CT 相比，BC 患者中有 259 个差异表达蛋白（DEP；155 个上调蛋白和 104 个下调蛋白）。 BC 衍生的 uEV 中上调的蛋白质富含与癌症进展相关的途径（即细胞增殖、细胞存活、细胞周期、细胞迁移、碳水化合物代谢和血管生成）。此外，我们使用 UALCAN 进行基于网络的验证来验证上调 DEP 的表达。值得注意的是，结果表明，与正常样本相比，BC 中 155 个上调蛋白中有 6 个（POSTN、ATAD2、BCAS4、GSK3β、HK1 和 Ki-67）过表达。由于这六种蛋白质通常充当细胞增殖和进展的标志物，因此它们可能是 BC 筛查和诊断的潜在生物标志物。然而，这需要在更大的队列中进行验证。版权所有：© 2023 Jeanmard 等人。这是一篇根据知识共享署名许可条款分发的开放获取文章，允许在任何媒体上不受限制地使用、分发和复制，前提是注明原始作者和来源。

Urinary extracellular vesicles (uEVs) reflect the biological conditions of the producing cells. The protein profiling of uEVs allow us to better understand cancer progression in several cancers such as bladder cancer, prostate cancer and kidney cancer but has not been reported in breast cancer. We have, herein, aimed at quantifying the concentration and at generating the proteomic profile of uEVs in patients with breast cancer (BC) as compared to that of healthy controls (CT). Urine samples were collected from 29 CT and 47 patients with BC. uEVs were isolated by using differential ultracentrifugation, and were then characterized by Western blotting and transmission electron microscopy. Moreover, a nanoparticle tracking analysis was used in order to measure the concentration and the size distribution of urine particles and uEVs. The proteomic profiling of the uEVs was facilitated through LC-MS/MS. The uEV concentration was not significantly different between the assessed groups. The undertaken proteomic analysis revealed 15,473 and 11,278 proteins in the BC patients' group and the CT group, respectively. Furthermore, a heat map analysis revealed a differential protein expression, while a principal component analysis highlighted two clusters. The volcano plot indicated 259 differentially expressed proteins (DEPs; 155 up- and 104 down-regulated proteins) in patients with BC compared with CT. The up-regulated proteins from BC-derived uEVs were enriched in pathways related to cancer progression (i.e., cell proliferation, cell survival, cell cycle, cell migration, carbohydrate metabolism, and angiogenesis). Moreover, we verified the expression of the upregulated DEPs using UALCAN for web-based validation. Remarkably, the results indicated that 6 of 155 up-regulated proteins (POSTN, ATAD2, BCAS4, GSK3β, HK1, and Ki-67) were overexpressed in BC compared with normal samples. Since these six proteins often act as markers of cell proliferation and progression, they may be potential biomarkers for BC screening and diagnosis. However, this requires validation in larger cohorts.Copyright: © 2023 Jeanmard et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.