Abstract
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-4H-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 (R2 = 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 GUS-overexpressed 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.
摘要
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.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (81801858), China Postdoctoral Science Foundation (2018M641676), and the Applied Basic Research Program of Shanxi Province (201801D221408).
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Zhou X, Zhang G, Ding D and Feng G designed the study and wrote the manuscript; Zhou X and Zhang H supervised the project and analyzed the data; Gao Z and Zhang G carried out the synthesis; Hou J detected the enzyme activity in the human serum; Zhou X, Wang D and He M performed the remaining experiments. All authors contributed to the general discussion.
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Xin Zhou received his PhD degree from the College of Life Sciences, Nankai University, in 2017. He is currently an associate professor at the College of Medical Imaging and Key Laboratory of Cellular Physiology, Ministry of Education, Shanxi Medical University. His current research focuses on the design and synthesis of nanomaterials for biomedical imaging and disease treatment.
Deping Wang is a lecturer at the Department of Physiology and Key Laboratory of Cellular Physiology, Ministry of Education, Shanxi Medical University. She received her PhD degree from Tianjin University in 2017. Her research interests include the crystal structures of proteins and nanotoxicology.
Hui Zhang is currently a vice president of Shanxi Medical University and director of the Department of Radiology, the First Hospital of Shanxi Medical University. She received her PhD degree (2002) from Shanghai Medical College of Fudan University, under the direction of Prof. Kangrong Zhou. Her research interests focus on the imaging diagnosis of central nervous system diseases, multimodal function magnetic resonance imaging, imaging genomics, and molecular imaging.
Guoqiang Zhang received his PhD degree from the College of Pharmacy, Nankai University in 2018. He is currently a postdoctoral fellow under the supervision of Prof. Dan Ding at the State Key Laboratory of Medicinal Chemical Biology, Nankai University. His current research focuses on the design and synthesis of smart/functional molecular imaging probes and exploration of their biomedical applications.
Guangxue Feng received his PhD degree from the Department of Chemical and Biomolecular Engineering, National University of Singapore in 2016. He is currently a professor at the State Key Laboratory of Luminescent Materials and Devices, South China University of Technology. His current research focuses on the design and synthesis of smart/functional optical nanomaterials for molecular imaging, phototheranostics and environment protection.
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Highly sensitive light-up near-infrared fluorescent probe for detection and imaging of β-glucuronidase in human serum, living cells and tumor-bearing mice
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Zhou, X., Wang, D., Gao, Z. et al. Highly sensitive light-up near-infrared fluorescent probe for detection and imaging of β-glucuronidase in human serum, living cells and tumor-bearing mice. Sci. China Mater. 65, 836–844 (2022). https://doi.org/10.1007/s40843-021-1779-7
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DOI: https://doi.org/10.1007/s40843-021-1779-7