Department of Urology, Osaka University Graduate School of Medicine, Yamadaoka, Suita, Osaka, 565-0871, Japan.
Department of Urology, Kindai University Faculty of Medicine, Ohno-Higashi, Osaka-Sayama, Osaka, 589-8511, Japan.
Br J Cancer. 2022 Oct;127(7):1312-1323. doi: 10.1038/s41416-022-01860-0. Epub 2022 Jul 6.
Urinary extracellular vesicles (uEVs) secreted from bladder cancer contain cancer-specific proteins that are potential diagnostic biomarkers. We identified and evaluated a uEV-based protein biomarker for bladder cancer diagnosis and analysed its functions.
Biomarker candidates, selected by shotgun proteomics, were validated using targeted proteomics of uEVs obtained from 49 patients with and 48 individuals without bladder cancer, including patients with non-malignant haematuria. We developed an enzyme-linked immunosorbent assay (ELISA) for quantifying the uEV protein biomarker without ultracentrifugation and evaluated urine samples from 36 patients with and 36 patients without bladder cancer.
Thirteen membrane proteins were significantly upregulated in the uEVs from patients with bladder cancer in shotgun proteomics. Among them, eight proteins were validated by target proteomics, and Ephrin type-A receptor 2 (EphA2) was the only protein significantly upregulated in the uEVs of patients with bladder cancer, compared with that of patients with non-malignant haematuria. The EV-EphA2-CD9 ELISA demonstrated good diagnostic performance (sensitivity: 61.1%, specificity: 97.2%). We showed that EphA2 promotes proliferation, invasion and migration and EV-EphA2 promotes the invasion and migration of bladder cancer cells.
We established EV-EphA2-CD9 ELISA for uEV-EphA2 detection for the non-invasive early clinical diagnosis of bladder cancer.
膀胱癌来源的尿细胞外囊泡（uEVs）中含有肿瘤特异性蛋白，是潜在的诊断生物标志物。我们鉴定并评估了基于 uEV 的膀胱癌诊断蛋白生物标志物，并分析了其功能。
采用鸟枪法蛋白质组学筛选候选生物标志物，并用靶向蛋白质组学检测 49 例膀胱癌患者和 48 例非膀胱癌个体（包括非恶性血尿患者）的 uEV 进行验证。我们开发了一种无需超速离心的 uEV 蛋白生物标志物酶联免疫吸附测定（ELISA），并评估了 36 例膀胱癌患者和 36 例非膀胱癌患者的尿液样本。
在鸟枪法蛋白质组学中，膀胱癌患者 uEV 中 13 种膜蛋白显著上调。其中，8 种蛋白经靶向蛋白质组学验证，EphA2 是膀胱癌患者 uEV 中唯一显著上调的蛋白，与非恶性血尿患者相比差异有统计学意义。EV-EphA2-CD9 ELISA 显示出良好的诊断性能（敏感性：61.1%，特异性：97.2%）。我们发现 EphA2 可促进增殖、侵袭和迁移，EV-EphA2 可促进膀胱癌细胞的侵袭和迁移。
我们建立了用于检测 uEV-EphA2 的 EV-EphA2-CD9 ELISA，用于非侵入性早期膀胱癌临床诊断。
