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Chitosan Combined with Molecular Beacon for Mir-155 Detection and Imaging in Lung Cancer

机译:壳聚糖结合分子信标在肺癌中Mir-155的检测和成像

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摘要

Lung cancer is the major cause of cancer-related deaths worldwide, thus developing effective methods for its early diagnosis is urgently needed. In recent years, microRNAs (miRNAs, miR) have been reported to play important roles in carcinogenesis and have become potential biomarkers for cancer diagnosis and treatment. Molecular beacon (MB) technology is a universal technology to detect DNA/RNA expression in living cells. As a natural polymers, chitosan (CS) nanoparticles could be used as a carrier for safe delivery of nucleic acid. In this study, we developed a probe using nanoparticles of miR-155 MB self assembled with CS (CS-miR-155 MB) to image the expression of miR-155 in cancer cells. Hybridization assay showed that the locked nucleic acid (LAN) modified miR-155 MB could target miR-155 effectively and sensitively. The miR-155 MB self-assembly with CS nanoparticles formed stable complexes at the proper weight ratio. The CS nanoparticles showed higher fluorescence intensity and transfection efficiency than the lipid-based formulation transfection agent by confocal microscopy and flow cytometry analysis. The CS-MB complexes were found to be easily synthesized and exhibited strong enzymatic stability, efficient cellular uptake, high target selectivity and biocompatibility. The CS-MB complexes can also be applied in other cancers just by simply changing for a targeted miRNA highly expressed in those cancer cells. Therefore, it is a promising vehicle used for detecting miRNA expression in living cells.
机译:肺癌是全世界与癌症相关的死亡的主要原因,因此迫切需要开发有效的方法对其进行早期诊断。近年来,据报道,微小RNA(miRNA,miR)在致癌作用中起着重要作用,并已成为癌症诊断和治疗的潜在生物标记。分子信标(MB)技术是检测活细胞中DNA / RNA表达的通用技术。作为天然聚合物,壳聚糖(CS)纳米粒子可用作安全递送核酸的载体。在这项研究中,我们开发了一种探针,该探针使用与CS自组装的miR-155 MB纳米颗粒(CS-miR-155 MB)来成像miR-155在癌细胞中的表达。杂交实验表明,锁定核酸(LAN)修饰的miR-155 MB可以有效,敏感地靶向miR-155。具有CS纳米粒子的miR-155 MB自组装以适当的重量比形成了稳定的复合物。通过共聚焦显微镜和流式细胞术分析,CS纳米颗粒显示出比基于脂质的制剂转染剂更高的荧光强度和转染效率。发现CS-MB复合物易于合成并显示出强的酶稳定性,有效的细胞摄取,高的靶标选择性和生物相容性。仅通过更改在那些癌细胞中高度表达的靶向miRNA,CS-MB复合物也可以应用于其他癌症。因此,它是用于检测活细胞中miRNA表达的有前途的载体。

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