基于功能化纳米探针与DNA自组装技术的肿瘤细胞荧光成像研究

In this work, a series of novel assay systems were developed for ultrasensitive fluorescence detection of cancer cell. The main researches include the design of novel, functional fluorescent nanoprobes and the study of self-assemble DNA strategy based on aptamer recognition. The high amplification efficiency could be produced by nanoprobe enhancement and self-assemble DNA amplification strategy. Based on the interaction of cancer cell and functional fluorescent nanoprobes, a series of detection method for cancer cell will be developed in this project by using laser confocal microscope and total internal reflection fluorescence microscope. The different markers expressed of the cancer cell will be detected using multiple fluorescent nanoprobes. The proposed methods may hold great promise for the broad applications in the field of medical research and early cancer diagnosis.

本项目拟开展肿瘤细胞及细胞内靶分子的荧光成像分析研究。通过制备新型的、多功能的、具有良好生物相容性的系列纳米荧光探针，提高细胞检测的准确性和灵敏度，结合核酸适体识别技术与DNA自组装信号放大技术，实现肿瘤细胞的原位检测与成像分析。利用激光共聚焦、全内反射等技术检测手段研究多功能纳米荧光探针与肿瘤细胞的相互作用过程，发展肿瘤细胞的高灵敏检测新方法，为荧光光谱在生物医学领域的应用提供重要参考与依据，为癌症的早期诊断提供新方法，新技术。

本项目以肿瘤细胞及肿瘤标志物为研究对象，将DNA生物传感与多种分析检测方法相结合，构建多种传感检测系统，实现多种肿瘤标志物的高灵敏检测。利用DNA自组装技术在细胞内形成纳米刷，实现细胞内miRNA及抑制肿瘤表达基因的成像及检测。将等温循环放大技术与阳离子交换，DNA分子步行器相结合，构建了多种传感检测系统实现对miRNA、DNA、T4PNK、外泌体等高灵敏检测。在此基础上，制备了AIE探针，利用静电作用及酶辅助信号放大技术，实现了对蛋白激酶CKII及DNA甲基化酶的活性检测。构建的多种传感体系对肿瘤标志物的检测进行了细致的研究，为后续课题的开展打下良好的基础。