Abstract
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.
Highlights
Xiaowei Cao, abce Shuai Chen,acd Zhenyu Wang,ac Yong Liu,f Xiaowei Luan,f Sicong Hou,abc Wei Li*ad and Hongcan Shi*abd
The results demonstrated that CYP2C9 with catalytic activity was expressed in 293T-Mig-2C9 cells, which provided a model for CYP2C9 function investigation
The successful synthesis of closely packed and long AuNPs–AgNWs in high yields was achieved by a facile electrostatic interaction method
Summary
Xiaowei Cao, abce Shuai Chen,acd Zhenyu Wang,ac Yong Liu,f Xiaowei Luan,f Sicong Hou,abc Wei Li*ad and Hongcan Shi*abd. 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 CYP2C9expressing 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 They were employed to fabricate SERS substrates via an electrostatically assisted 3-aminopropyltriethoxysilane (APTES)-functionalized surface-assembly method. SERS detection based on the AuNPs–AgNWs substrates provides a sensitive, nondestructive and label-free method for differentiation between 293T-Mig-R1 and 293T-Mig-2C9 cells. SERS has lots of advantages, for example, being nondestructive, showing weak water scattering and involving minimal sample preparation compared with other analytical techniques.[12] it is a powerful tool for studying living cells in systems containing water. AuNPs–AgNWs SERS substrates featuring rapid, label-free, and high sensitivity could serve as a promising tool for cell research, and the method shows potential for applications in high-throughput analysis, drug screening, and cell physiological process monitoring
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