MiR-145 detection in urinary extracellular vesicles increase diagnostic efficiency of prostate cancer based on hydrostatic filtration dialysis method
MiR-145 detection in urinary extracellular vesicles increase diagnostic efficiency of prostate cancer based on hydrostatic filtration dialysis method
复制标题
尿液细胞外囊泡中的 MiR-145 检测提高基于静水压滤过透析法的前列腺癌诊断效率
DOI：
10.1002/pros.23376
复制
发表时间：
2017-07-01
期刊：
影响因子：
2.8
通讯作者：
Zheng, Lei
中科院分区：
文献类型：
--
作者：
Xu, Yong;Qin, Sihua;Zheng, Lei
背景细胞外囊泡（Extracellular vesicles，EVs）可在体液中检测到，并可作为疾病的生物标志物.越来越多的证据表明，血清和尿液中循环的miRNA可能是前列腺癌（PCa）的潜在非侵入性生物标志物。在本研究中，我们的目的是调查是否静水过滤透析（HFD）是适合于尿EV（UEVs）的分离，以及是否可以检测到UEVs作为PCa生物标志物的PCa相关的miRNA。MethodsTo分析EVs的miRNA，我们寻找一种简单，经济的方法，以丰富EV从尿液样本。我们比较了HFD法和常规超离心法分离UEV的效率。随后，从PCa患者、良性前列腺增生（BPH）患者和健康个体中分离UEV。采用基于SYBR Green的定量逆转录-聚合酶链反应（qRT-PCR）检测UEV和配对血清EV中4种PCA相关miRNA（miR-572、miR-1290、miR-141和miR-145）的差异表达。在miRNA产量上，HFD和UC均能满足进一步分析的需要。前列腺癌患者UEVs中miR-145的表达水平显著高于BPH患者（P=0.018）。此外，与Gleason评分7相比，Gleason评分8肿瘤患者的miR-145水平显著增加（P=0.020）。受试者工作特征曲线（ROC）显示，UEV中的miR-145与血清PSA联合使用比PSA单独使用更能区分PCa和BPH（AUC分别为0.863和0.805）。在血清EV中，PCa患者中的四种miRNA显着高于BPH患者。结论与传统UC相比，HFD适用于UEV分离和miRNA分析。与BPH患者和健康对照相比，UEV中的miR-145在PCa患者中上调。我们建议UEVs miR-145作为PCa的生物标志物的潜在用途。
BackgroundExtracellular vesicles (EVs) can be detected in body fluids and may serve as disease biomarkers. Increasing evidence suggests that circulating miRNAs in serum and urine may be potential non-invasive biomarkers for prostate cancer (PCa). In the present study, we aimed to investigate whether hydrostatic filtration dialysis (HFD) is suitable for urinary EVs (UEVs) isolation and whether such reported PCa-related miRNAs can be detected in UEVs as PCa biomarkers.MethodsTo analyze EVs miRNAs, we searched for an easy and economic method to enrich EVs from urine samples. We compared the efficiency of HFD method and conventional ultracentrifugation (UC) in isolating UEVs. Subsequently, UEVs were isolated from patients with PCa, patients with benign prostate hyperplasia (BPH) and healthy individuals. Differential expression of four PCa-related miRNAs (miR-572, miR-1290, miR-141, and miR-145) were measured in UEVs and paired serum EVs using SYBR Green-based quantitative reverse transcription-polymerase chain reaction (qRT-PCR).ResultsThe overall performance of HFD was similar to UC. In miRNA yield, both HFD and UC can meet the needs of further analysis. The level of miR-145 in UEVs was significantly increased in patients with PCa compared with the patients with BPH (P=0.018). In addition, significant increase was observed in miR-145 levels when patients with Gleason score 8 tumors compared with Gleason score 7 (P=0.020). Receiver-operating characteristic curve (ROC) revealed that miR-145 in UEVs combined with serum PSA could differentiate PCa from BPH better than PSA alone (AUC 0.863 and AUC 0.805, respectively). In serum EVs, four miRNAs were significantly higher in patients with PCa than with BPH.ConclusionHFD is appropriate for UEVs isolation and miRNA analysis when compared with conventional UC. miR-145 in UEVs is upregulated from PCa patients compared BPH patients and healthy controls. We suggest the potential use of UEVs miR-145 as a biomarker of PCa.