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新型纳米 AS1411-壳聚糖-BODIPY 缀合物用于分子荧光成像。

Novel nanosized AS1411-chitosan-BODIPY conjugate for molecular fluorescent imaging.

机构信息

Department of Radiopharmacy, International Campus, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Int J Nanomedicine. 2019 May 15;14:3543-3555. doi: 10.2147/IJN.S202561. eCollection 2019.

Abstract

In recent years, non-invasive imaging technologies for early cancer detection have drawn worldwide attention. In this study, an antinucleolin aptamer, AS1411, was successfully conjugated to BODIPY-labeled chitosan and studied on T47D and HEK-293 cell lines. After conjugation of the aptamer to chitosan nanoparticles and purification, its structure was confirmed using atomic force microscopy (AFM), electrophoretic light scattering (ELS) and dynamic light scattering (DLS). Results of AFM, DLS and ELS of both conjugation and chitosan were compared for confirmation of conjugation. Conjugates were mixed with BODIPY FL fluorescent dye, purified and lyophilized. The labeled conjugate was characterized using Fourier-transform infrared spectroscopy, ultraviolet-visible spectroscopy, ELS and DLS. In vitro cellular uptake and cytotoxic effects of BODIPY-labeled chitosan-AS1411 aptamer conjugates were evaluated using the XTT assay on T47D and HEK-293 cells and flow cytometry on T47D cells. The data showed that uptake of BODIPY-labeled chitosan-AS1411 aptamer conjugate was satisfactory. Moreover, there was no statistically significant cytotoxicity of the conjugate on either cell line. The outcomes confirmed the potential application of this new targeted imaging agent as a novel cancer diagnostic agent for molecular imaging.

摘要

近年来,用于早期癌症检测的非侵入性成像技术引起了全球关注。在这项研究中,成功地将抗核仁素适体 AS1411 与 BODIPY 标记的壳聚糖缀合,并在 T47D 和 HEK-293 细胞系上进行了研究。 在将适体缀合到壳聚糖纳米粒子并进行纯化后,使用原子力显微镜(AFM)、电泳光散射(ELS)和动态光散射(DLS)确认其结构。比较了缀合前后的 AFM、DLS 和 ELS 结果,以确认缀合。将缀合物与 BODIPY FL 荧光染料混合、纯化和冻干。使用傅里叶变换红外光谱、紫外-可见光谱、ELS 和 DLS 对标记的缀合物进行了表征。使用 T47D 和 HEK-293 细胞的 XTT 测定法和 T47D 细胞的流式细胞术评估了 BODIPY 标记的壳聚糖-AS1411 适体缀合物的体外细胞摄取和细胞毒性作用。数据表明,BODIPY 标记的壳聚糖-AS1411 适体缀合物的摄取令人满意。此外,该缀合物在两种细胞系上均无统计学意义的细胞毒性。研究结果证实了这种新型靶向成像剂作为一种新型癌症诊断剂在分子成像中的潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7739/6526928/2c72da975875/IJN-14-3543-g0001.jpg

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