基于寡核酸探针信号放大microRNA活细胞内实时、动态荧光成像研究

项目名称： 基于寡核酸探针信号放大microRNA活细胞内实时、动态荧光成像研究

项目编号： No.21505052

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 刘海云

作者单位： 济南大学

项目金额： 21万元

中文摘要： 癌症的早期诊断与治疗已成为全球的关注热点和攻克难关。目前最新研究发现microRNA（miRNA）能够作为很好的癌症早期肿瘤标志物，但其在细胞内含量低、成像难。通常寡核酸探针进入细胞难度大，且进入细胞内的核酸探针在无DNA酶参与时往往不能实现细胞内miRNA信号级联放大。本项目利用DNA碱基Watson-Crick配对原理和DNA-石墨烯共轭堆积作用探索实现活细胞内miRNA信号级联放大和实时、动态成像方法。设计完成无DNA酶参与时核酸探针的跨膜及细胞内核酸扩增，以开发简单、有效监控活细胞内miRNA生物学过程的方法。通过miRNA上下游肿瘤标志蛋白、p53等的动态调节，观察miRNA在细胞内的网络信号调控和行为过程，探讨miRNA在肿瘤早期调控中的作用。期望基于此项目研究能够使癌症的早期发现和早期诊断获得进展，对于深入探讨生命现象的本质、解释细胞行为和疾病发生机制都具有重大意义。

中文关键词： 寡核酸探针；信号放大；MiroRNA；荧光检测；活细胞成像

英文摘要： The early diagnosis and treatment of cancer have become harsh difficulties and have gained global attention. Recently, microRNA (miRNA) is shown to be a good early stage tumor markers, but it is low levels in cells and imaging difficulty. In general, Oligonucleotide probes are difficult to be permeable into cells membrane, moreover, the probes cannot achieve signaling cascade amplification without DNA enzymes in living cells. The project will explores signaling cascade amplification and real-time dynamic imaging method of miRNA in living cells with Watson-Crick base-pairing principle and DNA-graphene stacking interactions. It will be achieve probes permeable into the cell membrane and amplification reaction in cell. We hope that monitor the biological processes of miRNA with a simple and effective method in living cells. To investigate the role of miRNA in the regulation of early stage cancer, we will observe the regulatory network signals and behavioral processes of miRNA in the cell, explore the dynamic adjustment of tumor marker in miRNA signaling network, such as protein, gene and p53. We expect it that the progress in the early detection and diagnosis of cancer. It can contribute to the further understanding of life phenomena, cell behavior and disease occurs.

英文关键词： Oligonucleotide Probes;Signal Amplification;MiroRNA;Fluorescence Detection;Cell Imaging