Weng Wen-Hui, Chiou Yueh-Er, Zeng Wen-Zhen, Chen Ming-Hui, Pang See-Tong
Department of Chemical Engineering and Biotechnology and Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei City, Taiwan.
Department of Nursing, College of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan.
Int J Nanomedicine. 2025 Jul 22;20:9291-9300. doi: 10.2147/IJN.S503954. eCollection 2025.
MicroRNAs (miRNAs) are small, non-coding RNA molecules critical for cellular function, growth, and development. Recent advances in remote diagnostic technologies have highlighted the potential of urinary miRNAs as non-invasive biomarkers for disease monitoring. This study introduces a simple, rapid, and cost-effective reagent for exosomal miRNA extraction, designed for urine-based exosome screening. We further identified hsa-miR-16-5p (miR-16) as a promising diagnostic biomarker for prostate cancer (PCa), with the goal of integrating this method with biosensors to enable rapid result acquisition and support early cancer detection and treatment planning.
The extraction reagent was formulated using polyethylene glycol (PEG), sodium dodecyl sulfate (SDS), and diethyl pyrocarbonate (DEPC), differing from commercial kits. miRNAs were extracted and validated through RT-qPCR, focusing on miR-16 and the reference control miR-21. Results were compared against commercial kits. A cohort of 39 clinical samples-28 PCa patients, 1 benign prostatic hyperplasia (BPH) case, and 10 healthy controls (Ctr)-was analyzed. Statistical evaluations included -tests and receiver operating characteristic (ROC) analysis to assess the diagnostic value of miR-16.
miR-16 expression significantly differed across PCa stages and between cancerous and non-cancerous individuals (Ctr vs Stage IV: < 0.05; Stage II vs III: < 0.05; Stage II vs IV: < 0.005; Stage III vs IV: < 0.05). ROC analysis confirmed miR-16's diagnostic potential, particularly in detecting mid-stage PCa.
The proposed extraction method matches commercial kits in performance but offers notable advantages: a simplified five-step process, reduced extraction time (1.08 hours), and low cost ($0.13/test). These findings support the use of urinary miR-16 as a biomarker for PCa staging and highlight the practical value of the newly developed extraction reagent.
微小RNA（miRNA）是对细胞功能、生长和发育至关重要的小的非编码RNA分子。远程诊断技术的最新进展凸显了尿miRNA作为疾病监测的非侵入性生物标志物的潜力。本研究介绍了一种用于外泌体miRNA提取的简单、快速且经济高效的试剂，专为基于尿液的外泌体筛查而设计。我们进一步将人miR-16-5p（miR-16）鉴定为前列腺癌（PCa）的一种有前景的诊断生物标志物，目标是将该方法与生物传感器整合，以实现快速获取结果并支持早期癌症检测和治疗规划。
提取试剂由聚乙二醇（PEG）、十二烷基硫酸钠（SDS）和焦碳酸二乙酯（DEPC）配制而成，与商业试剂盒不同。通过逆转录定量聚合酶链反应（RT-qPCR）提取并验证miRNA，重点关注miR-16和作为参考对照的miR-21。将结果与商业试剂盒进行比较。分析了一组39份临床样本，包括28例PCa患者、1例良性前列腺增生（BPH）病例和10例健康对照（Ctr）。统计评估包括t检验和受试者操作特征（ROC）分析，以评估miR-16的诊断价值。
miR-16的表达在PCa的不同阶段以及癌性和非癌性个体之间（Ctr与IV期：<0.05；II期与III期：<0.05；II期与IV期：<0.005；III期与IV期：<0.05）存在显著差异。ROC分析证实了miR-16的诊断潜力，尤其是在检测中期PCa方面。
所提出的提取方法在性能上与商业试剂盒相当，但具有显著优势：简化为五步流程、提取时间缩短（1.08小时）且成本低（0.13美元/测试）。这些发现支持将尿miR-16用作PCa分期的生物标志物，并突出了新开发的提取试剂的实用价值。
