拉曼响应的分子印记单层传感器对潜在肿瘤标志物金属硫蛋白的特异性识别和快速检测

Metallothioneins are a class of endogenous metal-binding proteins, whose content is closely correlated with the oncogenesis, development and chemoresistance of tumor cells. Therefore, it has scientific significance to monitor the concentration of metallothioneins from body for revealing various physiological phenomena and developing new methods of medical examination. In this project, the selectivity of molecularly imprinted materials and sensitivity of surface-enhanced Raman spectroscopy (SERS) are combined to design and prepare Raman-responded molecular imprinting monolayer sensor for direct identification and rapid detection of metallothionein from complex samples. Major research contents include: (1) We will explore the method of controllable growth of ordered core-shell metal nanoarray to prepare the SERS substrate with high sensitivity and good reproducibility; (2) We will develop the principle of molecular imprinting monolayer based on self-assembly to construct Raman-responded molecular imprinting monolayer sensor, and investigate the recognition process and enhancement mechanism of the obtained sensor; (3) We will establish the technology platform for the separation and analysis of metallothionein from tumor tissue to achieve rapid discrimination and detection. The expected results of this project will provide scientific basis and new approach for the quick monitor of potential tumor biomarker from clinical samples.

金属硫蛋白是一类内源性金属结合蛋白，其含量变化与肿瘤的发生、发展及耐药有明显相关性。因此，监测其在人体内的含量对于揭示各种生理现象及发展新的医学检验方法具有重要的科学意义。本项目将分子印记材料选择性与表面增强拉曼光谱(SERS) 敏感性相结合，设计和制备具有拉曼响应的分子印记单层传感器，实现复杂体系中金属硫蛋白的特异性识别和快速检测，研究内容如下：(1)探索有序芯-壳型金属纳米阵列的可控生长方法，制备出灵敏性高和重现性好的SERS衬底；(2)发展基于自组装的分子印记单层的原理，构建具有拉曼响应的分子印记单层传感器，并研究其对临床样本中金属硫蛋白的识别机制与拉曼信号放大输出的规律；(3)建立人体肿瘤组织中金属硫蛋白分离分析的技术平台，实现对金属硫蛋白的快速甄别与检测。本项目的预期成果将为临床样本中潜在肿瘤标志物的快速监测提供科学依据和新途径。

金属硫蛋白(MT)具有高诱导特性和多种生物学功能，广泛参与肿瘤细胞的分化、增殖和凋亡，以及肿瘤组织对顺铂类化疗药物的耐药。与正常组织相比，肿瘤组织中MT表达会发生一定程度上调或下调，其含量变化与肿瘤类型和分化程度具有明显相关性。因此，有效监测肿瘤组织中MT含量变化不仅可以提示肿瘤细胞的增殖活性，还可以提供预后相关信息，同时也可反映肿瘤细胞对化疗药物的耐受程度，对于揭示各种生理现象及发展新的医学检验方法具有重要的科学意义。本项目将拉曼光谱和分子印记技术相结合，在制备琼脂稳定的银包金纳米粒子凝胶膜的基础上，利用链状巯基烷烃和对巯基苯胺分别作为印记层分子和探针分子，通过对制备条件的优化，最终构建得到具有选择性识别能力的分子印记单层传感器，并用于MT检测，检测限可达1 μg/mL，相关系数为0.986.该方法具有操作简单、抗干扰能力强、样品用量少等优点，为发展基于分子印记的拉曼传感器以及监测临床样本中潜在肿瘤标志物提供了理论基础和新的途径。