针对醌氧化还原酶的荧光探针设计与活体癌细胞成像研究

Tumor is one of the major diseases threatening human health. For early warning, accurate diagnosis of tumors, accurate treatment, antitumor drug screening and evaluation of prognosis, it has the vital significance to develop a method for the high selective and sensitive detection of tumor markers in vivo. Quinone oxidoreductase 1 (NQO1), as a kind of important biological enzyme, has been confirmed to have close relationship with the occurrence and development of a variety of cancer cells in the organism. Aimed for the detection of NQO1 enzyme, the tumor marker related to tumor, a series of fluorescent probes with good biological compatibility and specific recognition ability were designed and synthesized. Based on pyridine/quinoline hemicyanine dyes, the fluorescent probes were constructed and derivated. And the enzyme content was determinated according to the fluorescence changes before and after the interaction between the probe molecule and the active site of enzyme. After completing the qualitative and quantitative detection of NQO1 enzyme in vitro, this project was aimed at further developing the fluorescence imaging technique for NQO1 enzyme detection and in vivo tracking of intracellular enzyme in the lesion site. At the same time, the technology was applied for the fluorescence detection of cancer patients' blood, tissue fluid, urine and other biological samples. Using this technology, a kind of rapid, high sensitive diagnostic kit was hopeful to be developed for early diagnosis of cancer.

肿瘤是威胁人类健康的重大疾病之一。发展针对生物体内肿瘤标志物的高选择性和高灵敏度的识别与检测方法，对于肿瘤的早期预警、准确诊断、精确治疗、抗肿瘤药物的筛选和预后评估等具有十分重要的意义。醌氧化还原酶1（NQO1），作为一种重要的生物酶类，被证实与生物体内多种癌细胞的发生与发展有着密切关系。本项目以 NQO1 酶这一与肿瘤密切相关的肿瘤标志物为研究对象，设计合成系列对该酶有特异性识别能力的、具有良好生物相容性的荧光探针分子。该荧光探针以吡啶/喹啉半菁类荧光染料作为母体结构，将其衍生化，利用探针分子与酶的活性位点作用前后荧光变化来检测酶的含量水平。该项目在完成体外NQO1 酶的定性、定量检测基础上，进一步发展体内检测NQO1 酶的荧光成像技术，实现对病变部位细胞内酶的活体示踪。同时，将该技术应用于癌症患者血液、组织液、尿液等生物样品的荧光检测，以期研制快速、高灵敏、针对癌症早期诊断的试剂盒。

肿瘤是威胁人类健康的常见疾病，针对肿瘤标志物的检测与成像已成为目前生物传感领域的研究热点之一。其中，醌氧化还原酶1（NQO1）是一种与肿瘤密切相关的重要生物酶类，被证实与生物体内多种癌细胞的发生与发展有着密切关系。.本项目研究主要以 NQO1 酶这一肿瘤标志物为研究对象，设计合成系列对该酶有特异性识别能力的、具有良好生物相容性的荧光探针分子。研究工作以半菁类化合物作为荧光报告基团，通过在分子骨架中引入NQO1酶的识别作用单元，合成了一系列吡啶型和喹啉型半菁类荧光探针分子。通过高分辨质谱、核磁等方法对荧光探针分子的结构进行了表征。系统考察了荧光探针分子对NQO1酶的光化学传感性能，通过PH值和干扰实验研究，筛选并建立高灵敏、高选择性的荧光传感体系。研究了荧光探针分子对NQO1酶检测的反应时间、线性范围、检测限以及相关体系的作用机制。通过酶促动力学和高效液相色谱方法，研究了探针分子与NQO1酶相互作用的识别传感机制。筛选具有良好生物相容性的荧光探针分子，应用于不同细胞的荧光成像研究，利用激光扫描共聚焦显微活细胞成像技术研究了探针分子对各种癌细胞内目标分析物的荧光成像性能。本项目拓展了半菁类荧光化合物在生物酶传感领域的应用，为研制低成本、快速、高灵敏、高通量的NQO1酶检测试剂盒，以及进一步构建癌症早期诊断与预警的光化学传感检测平台提供了理论依据。