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Breast cancer plasma biopsy by in situ determination of exosomal microRNA-1246 with a molecular beacon
Analyst ( IF 3.6 ) Pub Date : 2021-2-12 , DOI: 10.1039/d0an02224a Yun Chen 1, 2, 3, 4, 5 , Ling-Yan Zhai 5, 6, 7 , Li-Min Zhang 1, 2, 3, 4, 5 , Xiao-Shan Ma 1, 2, 3, 4, 5 , Zheng Liu 1, 2, 3, 4, 5 , Min-Min Li 5, 8, 9, 10 , Jin-Xiang Chen 1, 2, 3, 4, 5 , Wen-Jun Duan 1, 2, 3, 4, 5
Analyst ( IF 3.6 ) Pub Date : 2021-2-12 , DOI: 10.1039/d0an02224a Yun Chen 1, 2, 3, 4, 5 , Ling-Yan Zhai 5, 6, 7 , Li-Min Zhang 1, 2, 3, 4, 5 , Xiao-Shan Ma 1, 2, 3, 4, 5 , Zheng Liu 1, 2, 3, 4, 5 , Min-Min Li 5, 8, 9, 10 , Jin-Xiang Chen 1, 2, 3, 4, 5 , Wen-Jun Duan 1, 2, 3, 4, 5
Affiliation
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Liquid biopsy is becoming an innovative tool in precision oncology owing to its noninvasive identification of biomarkers circulating in the body fluid at various time points for continuous and real-time analysis of disease progression. MicroRNAs in blood exosomes are identified as a new promising class of potential biomarkers for cancer diagnostics and prognostics. Conventional detection of blood exosomal microRNAs need multiple-step, complicated, costly, and time-consuming sample preparation of exosomes isolation and RNA extract, which affect the accuracy and reproducibility of analytical results. In this work, we set up an in situ quantitative analysis of human plasma exosomal miR-1246 by a probe of 2′-O-methyl and phosphorothioate modified molecular beacon. The probe has outstanding nuclease resistance in highly active RNase A/T1/I, which makes it stable for direct application in blood samples. With rapid rupture of exosomes membrane by Triton X-100, the probe can enter exosomes to specifically target miR-1246 exhibiting quantitative fluorescent signals. Using the output signals as a diagnostic marker, we differentiated 33 breast cancer patients from 37 healthy controls with 97.30% sensitivity and 93.94% specificity at the best cutoff. The blood biopsy is simple without extracting plasma exosomes and their nucleic acids content, time-saving in about 2 h of total analysis process, and microvolumes needed for plasma sample, suggesting its good potential to clinical application.
中文翻译：
通过分子信标原位测定外泌体microRNA-1246进行乳腺癌血浆活检
由于液体活检无创地识别了在各个时间点在体液中循环的生物标记物，用于疾病进展的连续和实时分析，液体活检正成为精确肿瘤学的一种创新工具。血液外泌体中的MicroRNA被鉴定为用于癌症诊断和预后的新的有前途的潜在生物标记物。血液外泌体microRNA的常规检测需要外泌体分离和RNA提取物的多步骤，复杂，昂贵且耗时的样品制备，这会影响分析结果的准确性和可重复性。在这项工作中，我们通过2'- O探针建立了人血浆外泌体miR-1246的原位定量分析-甲基和硫代磷酸酯修饰的分子信标。该探针在高活性RNase A / T1 / I中具有出色的核酸酶抗性，使其可直接用于血样中，因此非常稳定。随着Triton X-100迅速使外泌体膜破裂，探针可以进入外泌体以特异性靶向显示定量荧光信号的miR-1246。使用输出信号作为诊断标记，我们以最佳截止值将97例30％的敏感性和93.94％的特异性与37例健康对照区分开来。血液活检简单，无需提取血浆外泌体及其核酸含量，在整个分析过程中节省约2小时的时间，并节省了血浆样品所需的微量样品，表明其具有良好的临床应用潜力。
更新日期：2021-02-18
中文翻译：
通过分子信标原位测定外泌体microRNA-1246进行乳腺癌血浆活检
由于液体活检无创地识别了在各个时间点在体液中循环的生物标记物，用于疾病进展的连续和实时分析，液体活检正成为精确肿瘤学的一种创新工具。血液外泌体中的MicroRNA被鉴定为用于癌症诊断和预后的新的有前途的潜在生物标记物。血液外泌体microRNA的常规检测需要外泌体分离和RNA提取物的多步骤，复杂，昂贵且耗时的样品制备，这会影响分析结果的准确性和可重复性。在这项工作中，我们通过2'- O探针建立了人血浆外泌体miR-1246的原位定量分析-甲基和硫代磷酸酯修饰的分子信标。该探针在高活性RNase A / T1 / I中具有出色的核酸酶抗性，使其可直接用于血样中，因此非常稳定。随着Triton X-100迅速使外泌体膜破裂，探针可以进入外泌体以特异性靶向显示定量荧光信号的miR-1246。使用输出信号作为诊断标记，我们以最佳截止值将97例30％的敏感性和93.94％的特异性与37例健康对照区分开来。血液活检简单，无需提取血浆外泌体及其核酸含量，在整个分析过程中节省约2小时的时间，并节省了血浆样品所需的微量样品，表明其具有良好的临床应用潜力。
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