Hubei Key Laboratory of Bioinorganic Chemistry & Materia Medica, Key Laboratory of Material Chemistry for Energy Conversion and Storage, Ministry of Education, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology , Wuhan 430074, People's Republic of China.
Shenzhen Institute of Huazhong University of Science and Technology, Shenzhen 518000, People's Republic of China.
Anal Chem. 2017 Feb 7;89(3):2073-2079. doi: 10.1021/acs.analchem.6b04696. Epub 2017 Jan 27.
Detections of telomerase activity in vitro and in living cells are of great importance for clinical diagnosis of cancer. In this work, an AIEgens-based bioprobe with two fluorescent signals for enhanced monitor of extracellular and intracellular telomerase activity is designed. After addition of telomerase, two positively charged AIEgens (Silole-R and TPE-H) bind to quencher group labeled primer (QP) and the extension repeated units, leading enhancement of two telomerase-triggered fluorescent signals. Furthermore, by combination the wider linear range in vitro and lower background in living cells imaging, the bioprobe is used to detect telomerase extracted from various cell lines (MCF-7, HeLa, E-J, and HLF), 50 bladder cancer patients' urine samples, 10 normal people's urine samples, and also applied in mapping telomerase activity inside living cells (MCF-7, HeLa, MDA-MB-231, and HT1080). The results show that this well-designed strategy can successfully detect telomerase activity in vitro and in living cells with high sensitivity, indicating the potential application of this method in cancer cells bioimaging and clinical cancer diagnosis.
在体外和活细胞中检测端粒酶活性对于癌症的临床诊断具有重要意义。在这项工作中，设计了一种基于 AIEgen 的生物探针，具有两个荧光信号，用于增强对外源和细胞内端粒酶活性的监测。加入端粒酶后，两个带正电荷的 AIEgen（Silole-R 和 TPE-H）与标记淬灭基团的引物（QP）和延伸重复单元结合，导致两个端粒酶触发的荧光信号增强。此外，通过结合体外更宽的线性范围和活细胞成像中的低背景，该生物探针用于检测来自各种细胞系（MCF-7、HeLa、E-J 和 HLF）、50 名膀胱癌患者的尿液样本、10 名正常人的尿液样本中的端粒酶，也应用于活细胞内端粒酶活性的定位（MCF-7、HeLa、MDA-MB-231 和 HT1080）。结果表明，这种精心设计的策略可以成功地检测体外和活细胞中端粒酶的活性，具有高灵敏度，表明该方法在癌细胞生物成像和临床癌症诊断中的潜在应用。
