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用于微小RNA原位成像及癌细胞精确光动力治疗的内源性激活信号放大检测法的构建

Construction of an Endogenously Activated Signal Amplification Assay for In Situ Imaging of MicroRNA and Guided Precise Photodynamic Therapy for Cancer Cells.

作者信息

Tian Feng, Wang Qi, Jiang Chengfang, Li Mengmeng, Zhang Yuqi, Yu Jing, Li Fengyan, Yan Tao, Ren Linlin, Gai Zhengxiao, Zhang Shusheng, Song Xinyue

机构信息

Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers, College of Medicine, Linyi University, Linyi, 276005, P. R. China.

College of Chemical Engineering, Northwest Minzu University, Lanzhou, 730030, P. R. China.

出版信息

Anal Chem. 2023 Apr 4;95(13):5601-5609. doi: 10.1021/acs.analchem.2c05148. Epub 2023 Mar 24.

Abstract

The construction of a sensitive strategy for in situ visualizing and dynamic tracing intracellular microRNA is of great importance. Via the toehold-mediated strand displacement process, the catalytic hairpin assembly (CHA) could offer several hundreds-fold signal amplifications. However, the CHA may produce certain background interferences since microRNA may exist in normal cells. In this work, we constructed an endogenously and sequentially activated signal amplification strategy to provide the amplified dual-color fluorescence imaging of microRNA in living cancer cells, which was comprised of two successive reaction processes: the activation of the preprotective catalytic probe by the endogenous glutathione (GSH) and the subsequent catalytic hairpin assembly on the surface of the upconversion nanoprobe triggered by the specific microRNA. Since the concentration of GSH in cancer cells was much higher than that in normal cells and the extracellular environment, the activation of the designed nanoprobe could be controlled at the desirable site. With the merits of the endogenous initiation and selective activation, the designed nanoprobe could achieve the bioimaging of microRNA in living cancer cells with high precision and reliability. Furthermore, via the introduction of a photosensitizer molecule into the DNA strand, the designed nanoplatform could achieve the precise photodynamic therapy (PDT) for cancer cells and malignant tumors under the irradiation of the NIR laser. This work provided a new avenue to achieve the accurate imaging of intracellular microRNA and guided precise PDT, which would offer powerful hints to the early diagnosis and therapy of malignant tumors.

摘要

构建一种用于原位可视化和动态追踪细胞内微小RNA的灵敏策略具有重要意义。通过基于引发链的链置换过程,催化发夹组装(CHA)可实现数百倍的信号放大。然而,由于微小RNA可能存在于正常细胞中,CHA可能会产生一定的背景干扰。在这项工作中,我们构建了一种内源性和顺序激活的信号放大策略,以实现活癌细胞中微小RNA的放大双色荧光成像,该策略由两个连续的反应过程组成:内源性谷胱甘肽(GSH)激活预保护催化探针,以及随后由特定微小RNA触发的上转换纳米探针表面的催化发夹组装。由于癌细胞中GSH的浓度远高于正常细胞和细胞外环境中的浓度,所设计纳米探针的激活可在理想位点得到控制。凭借内源性起始和选择性激活的优点,所设计的纳米探针能够高精度、可靠地实现活癌细胞中微小RNA的生物成像。此外,通过将光敏剂分子引入DNA链,所设计的纳米平台能够在近红外激光照射下对癌细胞和恶性肿瘤实现精确的光动力疗法(PDT)。这项工作为实现细胞内微小RNA的精确成像和指导精确PDT提供了一条新途径,这将为恶性肿瘤的早期诊断和治疗提供有力线索。

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