Several microRNAs (miRNAs) have been identified as cell-free biomarkers for detecting renal cell carcinoma (RCC). Droplet digital polymerase chain reaction (ddPCR) is a unique technology for nucleic acid quantification. It has the potential for superior precision, reproducibility, and diagnostic performance in identifying circulating miRNA biomarkers compared to conventional quantitative real-time PCR (qRT-PCR). This study aims to evaluate the performance of ddPCR compared to qRT- PCR in identifying miRNA biomarkers that differentiate malignant from benign renal masses. Potential biomarkers of RCC were identified from a literature review. RNA was extracted from the plasma of 56 patients. All the samples underwent analysis via ddPCR as well as qRT-PCR, and expression levels were recorded for the following miRNAs: miR-93, -144, -210, -221, and -222. Tumors were grouped into low-grade ccRCC, high-grade ccRCC, papillary RCC, and benign masses (primarily angiomyolipoma). The miRNA miR-210 (p= 0.034) and the combination of miRs-210 and miR-222 (p= 0.003) were expressed at significantly higher rates among those with RCC than those with benign masses, as measured by ddPCR. Using the combination of miR-210 and miR-222, ddPCR identified significant differences between the subgroups: papillary RCC versus benign (p= 0.03), low-grade ccRCC versus benign (p= 0.026), and high-grade ccRCC versus benign (p= 0.002). The only significant difference between these subgroups using qRT-PCR was between high-grade ccRCC and benign (p= 0.045). All the AUCs were significant when comparing each RCC subgroup with benign for both PCR technologies. Using a combination of miR-210 and miR-222, ddPCR identified significant differences between benign and malignant renal masses that were not identified as significant by conventional qRT-PCR.
多项研究已证实多种微小核糖核酸（miRNA）可作为无细胞生物标志物用于肾细胞癌（RCC）的检测。微滴式数字聚合酶链反应（ddPCR）是一种独特的核酸定量技术，与传统定量实时聚合酶链反应（qRT-PCR）相比，在循环miRNA生物标志物检测方面可能具有更高的精密度、可重复性和诊断效能。本研究旨在评估ddPCR与qRT-PCR在鉴别良恶性肾肿瘤miRNA生物标志物方面的性能差异。通过文献综述筛选出RCC的潜在生物标志物，对56例患者血浆样本进行RNA提取，所有样本均采用ddPCR和qRT-PCR技术检测miR-93、-144、-210、-221及-222的表达水平。肿瘤样本分为低级别透明细胞肾细胞癌（ccRCC）、高级别ccRCC、乳头状肾细胞癌及良性肿块（主要为血管平滑肌脂肪瘤）四组。ddPCR检测结果显示，RCC患者中miR-210（p=0.034）及miR-210与miR-222联合标志物（p=0.003）的表达水平显著高于良性肿块组。通过miR-210与miR-222联合检测，ddPCR在亚组间发现显著差异：乳头状肾细胞癌与良性组（p=0.03）、低级别ccRCC与良性组（p=0.026）、高级别ccRCC与良性组（p=0.002）。而qRT-PCR仅检测到高级别ccRCC与良性组间存在显著差异（p=0.045）。两种PCR技术比较各RCC亚组与良性组时，所有受试者工作特征曲线下面积（AUC）均具有统计学意义。采用miR-210与miR-222联合检测策略，ddPCR能够识别传统qRT-PCR未能检出的良恶性肾肿瘤间显著差异。
肿瘤(癌症)患者之家