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首页> 外文期刊>RSC Advances >The label-free detection and distinction of CYP2C9-expressing and non-expressing cells by surface-enhanced Raman scattering substrates based on bimetallic AuNPs–AgNWs
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The label-free detection and distinction of CYP2C9-expressing and non-expressing cells by surface-enhanced Raman scattering substrates based on bimetallic AuNPs–AgNWs

机译：基于双金属AuNPs-AgNWs的表面增强拉曼散射底物对CYP2C9表达细胞和非表达细胞的无标记检测和区分

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Cytochrome P450 2C9 (CYP2C9) is capable of catalyzing the biotransformation of endogenous compounds in cells, indicating that this enzyme could change the intracellular environment and is related to the pathogenesis of diseases. Currently, it is still a challenge to study the differences in cellular components between CYP2C9-expressing and non-expressing cells. In this study, employing a Au nanoparticles–Ag nanowires (AuNPs–AgNWs) decorated silicon wafer as a novel non-destructive and label-free tool, we applied surface-enhanced Raman scattering (SERS) spectroscopy to detect and distinguish the cellular composition of CYP2C9-expressing cells (293T-Mig-2C9) and non-expressing cells (293T-Mig-R1). AgNWs with high surface roughness were formed by modification of AuNPs onto their surface by electrostatic interactions, which enabled them to exhibit greatly enhanced SERS ability. Then, they were employed to fabricate SERS substrates via an electrostatically assisted 3-aminopropyltriethoxysilane (APTES)-functionalized surface-assembly method. The SERS substrates exhibited high sensitivity with a detection limit of 1 × 10 ~(?9) M for 4-mercaptobenzoic acid (4-MBA). Meanwhile, the SERS substrates exhibited good uniformity and reproducibility. The cytotoxicity assay demonstrated that the SERS substrates displayed good biocompatibility with 293T cells. Before SERS measurements, CYP2C9 constantly expressed cells (293T-Mig-2C9 cells) and control cells (293T-Mig-R1 cells) were constructed. The expression of CYP2C9 and the catalytic activity in the cells were checked. Using the AuNPs–AgNWs substrates as a high-performance in vitro sensing platform allowed us to obtain fingerprint spectra of 293T-Mig-R1 and 293T-Mig-2C9 cells. The difference spectra between the two cell lines were studied to interpret the spectral differences and gain insight into the biochemical variations. Finally, principal component analysis (PCA) score plots of the SERS spectra were also used to better view the differences between the two cell lines. SERS detection based on the AuNPs–AgNWs substrates provides a sensitive, non-destructive and label-free method for differentiation between 293T-Mig-R1 and 293T-Mig-2C9 cells.
机译：细胞色素P450 2C9（CYP2C9）能够催化细胞中内源性化合物的生物转化，表明该酶可以改变细胞内环境，并与疾病的发病机理有关。目前，研究表达CYP2C9的细胞与不表达CYP2C9的细胞之间的差异仍然是一个挑战。在这项研究中，我们采用Au纳米粒子-Ag纳米线（AuNPs-AgNWs）装饰的硅片作为一种新型的无损且无标签的工具，我们应用了表面增强拉曼散射（SERS）光谱技术来检测和区分细胞的组成。 CYP2C9表达细胞（293T-Mig-2C9）和非表达细胞（293T-Mig-R1）。具有高表面粗糙度的AgNW是通过静电相互作用将AuNPs修饰在其表面上而形成的，从而使它们表现出大大增强的SERS能力。然后，将它们用于通过静电辅助的3-氨丙基三乙氧基硅烷（APTES）功能化的表面组装方法制造SERS基板。 SERS底物显示出高灵敏度，对4-巯基苯甲酸（4-MBA）的检出限为1×10〜（？9）M。同时，SERS基板表现出良好的均匀性和可重复性。细胞毒性试验表明，SERS底物与293T细胞具有良好的生物相容性。在测量SERS之前，先构建CYP2C9恒定表达细胞（293T-Mig-2C9细胞）和对照细胞（293T-Mig-R1细胞）。检查细胞中CYP2C9的表达和催化活性。使用AuNPs-AgNWs底物作为高性能的体外传感平台，使我们能够获得293T-Mig-R1和293T-Mig-2C9细胞的指纹图谱。研究了两种细胞系之间的差异光谱，以解释光谱差异并深入了解生化变化。最后，SERS光谱的主成分分析（PCA）评分图还用于更好地查看两种细胞系之间的差异。基于AuNPs-AgNWs底物的SERS检测为293T-Mig-R1和293T-Mig-2C9细胞之间的分化提供了一种灵敏，无损且无标记的方法。

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