State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.
School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.
Mikrochim Acta. 2023 Mar 11;190(4):130. doi: 10.1007/s00604-023-05709-y.
A dual-mode nanoprobe was constructed to detect Bax messenger RNA (mRNA), consisting of gold nanotriangles (AuNTs), a Cy5-modified recognition sequence, and a thiol-modified DNA sequence. Bax mRNA is one of the key pro-apoptotic factors in the apoptosis pathway. Raman enhancement and fluorescence quenching of the signal group Cy5 were performed using AuNTs as substrates. The thiol-modified nucleic acid chain is partially complementary to the Cy5-modified nucleic acid chain to form a double strand and is linked to the AuNTs by the Au-S bond. When Bax mRNA is present, the Cy5-modified strand specifically binds to it to form a more stable duplex, making Cy5 far away from AuNTs, and SERS signal is weakened while fluorescence signal is enhanced. The nanoprobe can be used for the quantitative detection of Bax mRNA in vitro. Combined with the high sensitivity of SERS and the visualization of fluorescence, this method has good specificity and can be used for in situ imaging and dynamic monitoring of Bax mRNA during deoxynivalenol (DON) toxin-induced apoptosis of HepG2 cells. DON plays a pathogenic role mainly by inducing cell apoptosis. The results confirmed that the proposed dual-mode nanoprobe has good versatility in various human cell lines.
构建了一种双模式纳米探针来检测 Bax 信使 RNA（mRNA），由金纳米三角形（AuNTs）、Cy5 修饰的识别序列和巯基修饰的 DNA 序列组成。Bax mRNA 是细胞凋亡途径中关键的促凋亡因子之一。利用 AuNTs 作为基底，对信号基团 Cy5 进行 Raman 增强和荧光猝灭。巯基修饰的核酸链与 Cy5 修饰的核酸链部分互补形成双链，并通过 Au-S 键与 AuNTs 连接。当存在 Bax mRNA 时，Cy5 修饰的链特异性地与其结合形成更稳定的双链，使 Cy5 远离 AuNTs，SERS 信号减弱而荧光信号增强。该纳米探针可用于体外 Bax mRNA 的定量检测。结合 SERS 的高灵敏度和荧光的可视化，该方法具有良好的特异性，可用于 DON 毒素诱导 HepG2 细胞凋亡过程中 Bax mRNA 的原位成像和动态监测。DON 主要通过诱导细胞凋亡发挥致病作用。结果证实，所提出的双模式纳米探针在各种人类细胞系中具有良好的通用性。
