A Pilot Study on the Potential of RNA-Associated to Urinary Vesicles as a Suitable Non-Invasive Source for Diagnostic Purposes in Bladder Cancer

Abstract: Bladder cancer is one of the most common cancers and, together with prostate carcinoma, accounts for the majority of the malignancies of the genitourinary tract. Since prognosis ameliorates with early detection, it will be beneficial to have a repertoire of diagnostic markers that could complement the current diagnosis protocols. Recently, cell-secreted extracellular vesicles have received great interest as a source of low invasive disease biomarkers because they are found in many body fluids, including urine. The current work describes a pilot study to generate an array-based catalogue of mRNA associated to urinary vesicles, and also a comparison with samples obtained from bladder cancer patients. After an analysis of presence/absence of transcripts in bladder cancer EVs, a list of genes was selected for further validation using PCR technique. We found four genes differentially expressed in cancer samples. LASS2 and GALNT1 were present in cancer patients, while ARHGEF39 and FOXO3 were found only in non-cancer urinary vesicles. Previous studies have pointed to the involvement of those genes in tumour progression and metastasis. Total RNA isolated from Extracellular Vesicles obtained from urine of bladder cancer patients was compared with RNA isolated from urinary vesicles of non-cancer patients.

摘要：膀胱癌是最常见的恶性肿瘤之一，与前列腺癌一同构成了泌尿生殖道恶性肿瘤的绝大多数病例。由于早期检测可改善患者预后，因此开发可辅助现有诊断方案的诊断标志物库具有重要应用价值。近年来，细胞分泌的细胞外囊泡（extracellular vesicles, EVs）作为低侵入性疾病生物标志物的来源受到广泛关注，因为它们广泛存在于包括尿液在内的多种体液中。本研究描述了一项先导性研究：构建基于芯片的尿液囊泡相关信使RNA（messenger RNA, mRNA）目录，并将其与膀胱癌患者的样本进行对比分析。在分析膀胱癌细胞外囊泡的转录本存在情况后，我们筛选出一组基因，采用聚合酶链反应（polymerase chain reaction, PCR）技术进行后续验证。最终发现4个在癌样本与非癌样本中差异表达的基因：LASS2与GALNT1在癌症患者样本中存在，而ARHGEF39与FOXO3仅在非癌性尿液囊泡中被检出。既往研究已表明，上述基因均参与肿瘤进展与转移过程。本研究将从膀胱癌患者尿液中分离的细胞外囊泡总RNA，与非癌症受试者尿液囊泡的总RNA进行了对比分析。