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利用分子信标检测外泌体 miRNA 诊断前列腺癌。

Detection of exosome miRNAs using molecular beacons for diagnosing prostate cancer.

机构信息

a Division of Bioengineering , Incheon National University , Incheon , Republic of Korea.

b Biomedical Omics Group , Korea Basic Science Institute , Cheongju , Republic of Korea.

出版信息

Artif Cells Nanomed Biotechnol. 2018;46(sup3):S52-S63. doi: 10.1080/21691401.2018.1489263. Epub 2018 Jul 22.

Abstract

Prostate cancer is the fifth leading cause of cancer-related deaths among males worldwide. However, the biomarker for diagnosing prostate cancer that is used currently has limitations that must be overcome. Recently, several studies have demonstrated that the cancer liquid biopsy can be implemented by using exosome miRNAs. However, the current methods for the detection of exosome miRNAs are time-consuming, expensive, and laborious. Thus, we investigated a novel method for diagnosing prostate cancer that involves the use of molecular beacons for the in situ detection of miRNAs in exosomes from prostate cancer cells. We chose miRNA-375 and miRNA-574-3p as the target miRNAs for prostate cancer, and these markers in exosomes produced by prostate cancer cells including DU145 and PC-3 were successfully detected using molecular beacons. High fluorescent signals were obtained from MB and miRNA hybridization in exosomes in a concentration-dependent manner. In addition, exosome miRNAs can be detected even in the presence of human urine, so this method can be applied directly using human urine to perform liquid biopsies for prostate cancer. Overall, the in situ detection of exosome miRNAs using molecular beacons can be developed as a simple, cost effective, and non-invasive liquid biopsy for diagnosing prostate cancer.

摘要

前列腺癌是全球男性癌症相关死亡的第五大主要原因。然而,目前用于诊断前列腺癌的生物标志物存在必须克服的局限性。最近,几项研究表明,癌症液体活检可以通过使用外泌体 miRNA 来实现。然而,目前检测外泌体 miRNA 的方法既耗时、昂贵又繁琐。因此,我们研究了一种新的诊断前列腺癌的方法,该方法涉及使用分子信标原位检测前列腺癌细胞中外泌体中的 miRNA。我们选择 miRNA-375 和 miRNA-574-3p 作为前列腺癌的靶 miRNA,并且使用分子信标成功地检测了前列腺癌细胞(包括 DU145 和 PC-3)产生的外泌体中的这些标记物。荧光信号强度与外泌体中 miRNA 与分子信标的杂交浓度呈正相关。此外,即使存在人尿,也可以检测到外泌体 miRNA,因此该方法可以直接用人尿进行液体活检以诊断前列腺癌。总之,使用分子信标原位检测外泌体 miRNA 可以作为一种简单、经济有效且非侵入性的液体活检方法用于诊断前列腺癌。

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