Highly sensitive light-up near-infrared fluorescent probe for detection and imaging of β-glucuronidase in human serum,living cells and tumor-bearing mice  

用于血清、活细胞及小鼠荷瘤中β-葡萄糖醛酸苷酶检测成像的高灵敏点亮型近红外荧光探针

作　　者：Xin Zhou Deping Wang Zhiyuan Gao Mingyue He Jiayi Hou Hui Zhang Guoqiang Zhang Dan Ding Guangxue Feng 周鑫;王德平;高志远;何明玥;侯佳宜;张辉;张国强;丁丹;冯光雪(College of Medical Imaging,and Key Laboratory of Cellular Physiology,Ministry of Education,Shanxi Medical University,Taiyuan 030001,China;Key Laboratory of Bioactive Materials Ministry of Education,College of Life Sciences,and State Key Laboratory of Medicinal Chemical Biology,Nankai University,Tianjin 300071,China;Department of Clinical Laboratory,Shanxi Provincial Academy of Traditional Chinese Medicine,Taiyuan 030001,China;AIE Institute,State Key Laboratory of Luminescent Materials and Devices,Guangdong Provincial Key Laboratory of Luminescence from Molecular Aggregates,School of Materials Science and Engineering,South China University of Technology,Guangzhou 510640,China)

机构地区：[1]College of Medical Imaging,and Key Laboratory of Cellular Physiology,Ministry of Education,Shanxi Medical University,Taiyuan 030001,China [2]Key Laboratory of Bioactive Materials Ministry of Education,College of Life Sciences,and State Key Laboratory of Medicinal Chemical Biology,Nankai University,Tianjin 300071,China [3]Department of Clinical Laboratory,Shanxi Provincial Academy of Traditional Chinese Medicine,Taiyuan 030001,China [4]AIE Institute,State Key Laboratory of Luminescent Materials and Devices,Guangdong Provincial Key Laboratory of Luminescence from Molecular Aggregates,School of Materials Science and Engineering,South China University of Technology,Guangzhou 510640,China

出　　处：《Science China Materials》2022年第3期836-844,共9页中国科学（材料科学（英文版）

基　　金：supported by the National Natural Science Foundation of China(81801858);China Postdoctoral Science Foundation(2018M641676);the Applied Basic Research Program of Shanxi Province(201801D221408)。

摘　　要：β-Glucuronidase(GUS)plays a key role in tumor initiation,metastasis,and progression,and thus,has been proposed as a promising cancer biomarker.In this study,we designed an enzyme-activatable near-infrared(NIR)fluorescent probe(DCM-βGlcA)for the rapid and accurate detection of GUS activity in vitro,in vivo and ex vivo.The DCM-βGlcA was prepared by linking a glucuronic acid residue to dicyanomethylene-4 H-pyran(DCM).This probe exhibited significant light-up NIR fluorescent signals at 680 nm after reacting with GUS and the Stokes shift could reach 150 nm.The DCM-βGlcA showed a high sensitivity toward GUS and an excellent linear relationship at concentrations ranging between 0 and 4 U L^(-1)(R^(2)=0.9974)with the limit of detection as low as 0.19 U L^(-1).We used the DCM-βGlcA to identify GUS serum levels in both cancer patients and healthy individuals with a similar accuracy as that of an enzyme-linked immunosorbent assay(ELISA)while being easier and faster to perform.Moreover,the DCM-βGlcA was used for tracking endogenous GUS in living cells,thereby discriminating GUSoverexpressed liver cancer from normal cells.Additionally,the DCM-βGlcA was able to detect and image endogenous GUS in liver cancer tissue and tumor-bearing mouse models.These findings demonstrate the potential of the DCM-βGlcA as a promising tool for detecting and monitoring GUS activity in preclinical applications.β-葡萄糖醛酸苷酶(GUS)在肿瘤的发生、转移和进展中具有关键作用,因而成为一种很有应用前景的癌症生物标志物.在本研究中,我们设计出一种酶激活的近红外荧光探针(DCM-βGlcA),可在体内和体外快速、准确地检测GUS的活性.该探针是由葡萄糖醛酸残基与荧光基团吡喃腈(DCM)衍生物构成.在GUS作用下,该探针在680 nm处具有明显的近红外荧光增强,而且它的斯托克位移达到了150 nm.此外,该探针对于GUS有很高的灵敏度,检测限低至0.19 U L^(-1).我们利用该探针分析了人血清样本中的GUS含量,其结果与ELISA检测结果一致.该探针还可用于示踪活细胞中的GUS,而且能够对荷瘤小鼠体内和肝癌组织中的GUS进行检测成像.因此,该探针可作为检测GUS的理想工具.

关 键 词：NIR fluorescent probe Β-GLUCURONIDASE serum detection cell imaging in-vivo bioimaging 

分 类 号：R730.4[医药卫生—肿瘤] O657.3[医药卫生—临床医学]