模块化组装超分子荧光纳米探针的构建及其在人肺腺癌和炎症区分中的应用研究

Lung cancer ranks first in the incidence of malignant tumors, and early diagnosis of lung cancer is the key to timely treatment. The clinical pneumonia and lung cancer can easily be confused. Therefore, how to effectively identify pneumonia and lung cancer is of great significance to improve the survival of patients. Because of the signal enhancement effect, nanoprobes have been often used to improve the sensitivity of tumor detection, but they are not effective to identify tumor and inflammation. To address this problem, combining the aptamer that specifically recognize tumor cells with the folic acid that can target folate receptors of activated macrophages in inflammation, this project, using the human lung adenocarcinoma cell as a model, plans to construct modularly assembled supramolecular fluorescent nanoprobes based on host-guest inclusion interactions between adamantane derivatives of fluorescent dyes and β-cyclodextrin polymer for identification of human lung adenocarcinoma cells, activated macrophages and normal cells. On this basis, the nanoprobes will be further used to distinguish between human lung adenocarcinoma tissues and inflammatory tissues. Through the self-assembling approach, the project is expected to build a simple, sensitive, and specific labeling technique for effective identification of tumor and inflammation at the same time, which could provide a valuable detection method and technical support for the diagnosis of human lung adenocarcinoma.

肺癌居恶性肿瘤发病率的首位，早期诊断肺癌是及时治疗的关键。而临床上极易将肺炎和肺癌相混淆，因此有效地鉴别肺炎与肺癌对于提高患者生存期意义重大。纳米探针因具备信号增强效应常被用于提高对肿瘤检测的灵敏度，但尚不能有效鉴别肿瘤和炎症。针对这一问题，本项目拟以人肺腺癌细胞A549为研究模型，基于β-环糊精聚合物与客体分子（金刚烷衍生化的荧光染料分子）间的主客体相互作用，结合核酸适配体对肿瘤细胞的特异性识别及叶酸可靶向识别炎症反应时活化的巨噬细胞表面叶酸受体的性质，构建模块化组装超分子荧光纳米探针；在研究该类纳米探针对人肺腺癌细胞、活化的巨噬细胞和正常细胞识别的基础上，将其进一步用于对人肺腺癌组织和炎症组织的区分。研究项目预期基于该自组装方法构建一种简便的、灵敏的、特异性的，可同时区别炎症和癌症的标记技术，从而为人肺腺癌的诊断提供有价值的检测方法和技术支持。

基于自组装方法制备纳米探针具备简便的、可模块化的性能，以及选择性地联合运用靶向识别分子可改善方法特异性的优点，本项目构建了一系列具备特异性识别功能的模块化组装纳米探针用于肿瘤患者和正常人的区分。首先，制备了模块化组装聚辣根过氧化物酶探针，并将其与磁分离富集技术相结合，建立了一种高灵敏、高特异性的方法用于人血清中白血病相关生物标志物的检测，肿瘤患者组与正常人组检测结果之间的差异有统计学意义（P < 0.001），表明所建方法具备临床应用价值。其次，构建了模块化组装核酸适配体/蛋白质复合纳米探针，结合核酸适配体功能化的磁纳米颗粒，实现了全血中对白血病细胞的有效捕获和高灵敏、快速检测。最后，基于新发现的细胞辅助增强化学发光模式，结合化学发光增强的反应机理研究，实现了对肿瘤细胞的快速和免标记检测，白血病患者全血样品的检测信号与健康人全血样品的检测信号存在显著性差异，表明其具备潜在的临床应用价值。