The label-free detection and distinction of CYP2C9-expressing and non-expressing cells by surface-enhanced Raman scattering substrates based on bimetallic AuNPs-AgNWs

Xiaowei Cao; Shuai Chen; Zhenyu Wang; Yong Liu; Xiaowei Luan; Sicong Hou; Wei Li; Hongcan Shi

doi:10.1039/c9ra02046b

The label-free detection and distinction of CYP2C9-expressing and non-expressing cells by surface-enhanced Raman scattering substrates based on bimetallic AuNPs-AgNWs

RSC Adv. 2019 Apr 30;9(23):13304-13315. doi: 10.1039/c9ra02046b. eCollection 2019 Apr 25.

Authors

Xiaowei Cao^{1

2

3

4}, Shuai Chen^{1

3

5}, Zhenyu Wang^{1

3}, Yong Liu⁶, Xiaowei Luan⁶, Sicong Hou^{1

2

3}, Wei Li^{1

5}, Hongcan Shi^{1

2

5}

Affiliations

¹ Institute of Translational Medicine, Medical College, Yangzhou University Yangzhou 225001 PR China weili@yzu.edu.cn shihongcan@yzu.edu.cn.
² Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University Yangzhou 225001 PR China.
³ Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University Yangzhou 225009 China.
⁴ The Key Laboratory of Syndrome Differentiation and Treatment of Gastric Cancer of the State Administration of Traditional Chinese Medicine Yangzhou 225001 PR China.
⁵ Jiangsu Key Laboratory of Experimental & Translational Non-coding RNA Research Yangzhou 225001 PR China.
⁶ School of Life Science and Medicine, Dalian University of Technology Panjin 124221 China.

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.