Ministry of Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules & Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Hubei University , Wuhan 430062, P. R. China.
Key Laboratory for Green Chemical Process of Ministry of Education and School of Chemistry and Environmental Engineering, Wuhan Institute of Technology , Wuhan 430205, P. R. China.
Anal Chem. 2017 Nov 7;89(21):11576-11582. doi: 10.1021/acs.analchem.7b02910. Epub 2017 Oct 26.
Leucine aminopeptidase (LAP) is an important cancer-related biomarker, which shows significant overexpression in malignant tumor cells like liver cancer. Developing an effective method to monitor LAP in tumor cells holds great potential for cancer diagnosis, treatment, and management. In this work, we report a novel BODIPY-based fluorescent probe (BODIPY-C-Leu) capable of monitoring LAP in vitro and in vivo in both ratiometric and turn-on model. BODIPY-C-Leu contains an asymmetrical BODIPY dye for fluorescent signaling and a dipeptide (Cys-Leu) as the triggered moiety. Activation occurs by cleavage of the amide bond in dipeptides and subsequently an intramolecular S → N conversion to convert sulfur-substituted BODIPY to amino-substituted BODIPY, resulting in a dramatic fluorescence variation to realize the detection of LAP. Furthermore, we have successfully employed BODIPY-C-Leu to monitor LAP activity in different cancer cells, indicating that HeLa cells have a higher level of LAP activity than A549 cells. Importantly, we demonstrated the capability of the probe for real-time monitoring the drug-induced LAP level changes in zebrafish.
亮氨酸氨基肽酶（LAP）是一种重要的与癌症相关的生物标志物，在肝癌等恶性肿瘤细胞中表现出明显的过表达。开发一种有效监测肿瘤细胞中 LAP 的方法，对于癌症的诊断、治疗和管理具有巨大的潜力。在这项工作中，我们报告了一种新型基于 BODIPY 的荧光探针（BODIPY-C-Leu），能够在体外和体内以比率和开启两种模式监测 LAP。BODIPY-C-Leu 包含一个不对称的 BODIPY 染料用于荧光信号，以及一个二肽（Cys-Leu）作为触发部分。通过二肽中酰胺键的断裂和随后的分子内 S → N 转换，将硫取代的 BODIPY 转化为氨基取代的 BODIPY，从而导致荧光显著变化，实现 LAP 的检测。此外，我们已经成功地利用 BODIPY-C-Leu 来监测不同癌细胞中的 LAP 活性，表明 HeLa 细胞的 LAP 活性高于 A549 细胞。重要的是，我们证明了该探针实时监测斑马鱼中药物诱导的 LAP 水平变化的能力。
