基于SERS的肿瘤细胞外泌体及其内含RNA检测研究

In this work, surface enhanced Raman scattering (SERS) technique is used to detect tumor derived exosomes and exosomal RNAs. First, we will construct SERS nanoprobes, immunomagnetic nanobeads and SERS substrates which can recognize exosomal RNAs. Tumor derived exosomes will be identified via SERS signals of the nanoprobes, while exosomal RNAs will be analyzed via SERS and fluorescence signals of molecular beacons modified on the SERS substrates. Advantages of the proposed method include: 1) SERS technique can achieve an extremely high sensitivity, meanwhile, it can fulfill simultaneous detection of multiple kinds of exosomes and exosomal RNAs owing to its high optical encoding capacity, which is hard to achieve by other methods. 2) Body fluids will be used in the detection, which can greatly reduce physical injuries to the patients. Moreover, this will allow real-time monitoring of tumor progression and drug reactions, which will facilitate timely adjustment of tumor therapies. We will investigate the fabrication strategies of the SERS nanoprobes, immunomagnetic nanobeads and SERS substrates as well as the optimization of their performances. We will also study multiplex detection of tumor exosomes and exosomal RNAs. We have already made important preliminary progress in the fabrication of optical nanoprobes. The proposed work shows a vital significance and a promising clinical applicability in the early diagnosis of tumors, the investigation of tumor metastasis mechanisms and the therapeutics of multiple primary carcinomas.

本项目提出采用SERS光谱技术检测体液中的肿瘤细胞外泌体及内含RNA。项目将首先构建出SERS纳米探针、免疫磁球及能识别内含RNA的SERS基底。外泌体的种类通过纳米探针的SERS信号区分；内含RNA通过SERS基底上的分子信标的SERS及荧光信号分析。该检测方案具有以下优势：1）SERS光谱技术的引入，一方面可实现超高灵敏的检测，另一方面由于其光谱编码容量大，有望解决同时检测多种肿瘤细胞外泌体及内含RNA的难题；2）利用体液检测，能最大程度地降低对病人的生理损伤，同时还能实时监测肿瘤发展的状态及药物反应，方便及时调整治疗方案。项目将研究SERS纳米探针、免疫磁球和SERS基底的制备技术和性能优化方法，同时将研究外泌体及内含RNA的多元检测技术。项目在纳米探针合成方面已取得了重要的前期进展。项目的研究对肿瘤早期诊断、转移机理研究以及多原发癌患者的治疗具有重要的科学意义与实际临床应用价值。

本项目提出采用SERS光谱技术检测体液中的肿瘤细胞外泌体及内含RNA。项目首先构建出SERS纳米探针、免疫磁球及能识别内含RNA的SERS基底。外泌体的种类通过纳米探针的SERS信号区分；内含RNA通过SERS基底上的分子信标的SERS及荧光信号分析。项目研究了SERS纳米探针、免疫磁球和SERS基底的制备技术和性能优化方法，同时研究了外泌体及内含RNA的多元检测技术。通过三年的研究，成功实现了基于SERS的肿瘤细胞外泌体及内含RNA检测。研究中制备出了具有新型结构的SERS纳米探针粒子以及捕获基底，在原有申请书固相或者液相基底的基础上，升级为在微流控芯片中进行检测，大大提高了分析的效率。实验中能同时进行三种肿瘤细胞外泌体及内含RNA的检测，外泌体检测限比SPR技术提高近100倍。此外，本项目发展的技术还能用于真实血液样本中的外泌体检测，展现出很好的临床癌症筛查潜力。项目的研究对肿瘤早期诊断、转移机理研究以及多原发癌患者的治疗具有重要的科学意义与实际临床应用价值。