Cheng Lingling, Xu Jianlin, Yuan Hua, Zhao Qihao, Yue Wei, Ma Shuang, Lu Weimin
Department of Oncology, Yancheng TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Yancheng, Jiangsu, China.
Pharmacy Department, Yancheng TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Yancheng, Jiangsu, China.
Front Bioeng Biotechnol. 2023 Dec 7;11:1310258. doi: 10.3389/fbioe.2023.1310258. eCollection 2023.
In this study, a surface-enhanced Raman scattering (SERS) sensor based on a functionalized Au/Si cap-cone array (Au/Si CCA) was constructed using the identity-release strategy to detect Vimentin changes during epithelial-mesenchymal transition (EMT) in gastric cancer (GC). The periodic structure of Au/Si CCA, which can form "hot spots" with high density and regular arrangement, is a substrate with excellent performance. Au/Si CCA was functionalized with aptamers as the capture substrate, and Au nanocubes (AuNCs) were modified with 5-carboxyfluorescein (5-FAM) labelled complementary strand as SERS probe. The capture substrate and SERS probe were assembled by hybridization, and the SERS signal intensity of 5-FAM was greatly enhanced. The binding of Vimentin to the aptamer resulted in a broken connection between the SERS sensor Au/Si CCA array and AuNCs, which resulted in a decrease in the signal intensity of 5-FAM. The identity-release strategy requires only a simple step of reaction to achieve rapid detection of target proteins, which has clinical practicability. Using this protocol, the concentration of Vimentin in GES-1 cells could be successfully detected, and the detection limit was as low as 4.92 pg/mL. Biological experiments of Vincristine, Oncovin (VCR)-treated GES-1 cells effectively mimicked the EMT process, and Vimentin changes during EMT could be accurately detected by this method. This study provides a selective, ultra-sensitive and accurate assay for Vimentin detection, which may provide a means for the future detection of EMT process in GC.
在本研究中,基于功能化金/硅帽锥阵列(Au/Si CCA)构建了一种表面增强拉曼散射(SERS)传感器,采用身份释放策略来检测胃癌(GC)上皮-间质转化(EMT)过程中波形蛋白的变化。Au/Si CCA的周期性结构能够形成高密度且规则排列的“热点”,是一种性能优异的基底。以适配体对Au/Si CCA进行功能化作为捕获基底,用5-羧基荧光素(5-FAM)标记的互补链修饰金纳米立方体(AuNCs)作为SERS探针。捕获基底和SERS探针通过杂交组装,5-FAM的SERS信号强度得到极大增强。波形蛋白与适配体的结合导致SERS传感器Au/Si CCA阵列与AuNCs之间的连接断裂,从而导致5-FAM信号强度降低。身份释放策略仅需简单的一步反应即可实现对目标蛋白的快速检测,具有临床实用性。采用该方案能够成功检测GES-1细胞中波形蛋白的浓度,检测限低至4.92 pg/mL。长春新碱(Vincristine,Oncovin,VCR)处理GES-1细胞的生物学实验有效模拟了EMT过程,且该方法能够准确检测EMT过程中波形蛋白的变化。本研究为波形蛋白检测提供了一种选择性高、超灵敏且准确的分析方法,可能为未来检测GC中的EMT过程提供一种手段。