基于功能化高阶框架核酸的前列腺肿瘤多标志物联合检测研究

Prostate cancer is one of the most common malignant tumors, its accurate diagnosis has become the key to reduce the mortality and improve the prognosis of prostate cancer. As cancer occurrence is a complicated process driven by multiple genes and activated by multiple signal transduction pathways, its diagnostic accuracy based on joint detection of multiple relevant markers would be higher than that based on single marker detection. With delicate structure, the programmable framework nucleic acid has provided a new approach for biological interface regulation and highly sensitive biomolecular sensing. In this proposal, functional framework nucleic acid tetrahedron with specific response to a single tumor marker of prostatic cancer will be firstly constructed. Then higher-order framework nucleic acid structures with multiple integrated functions will be assembled by adopting the tetrahedrons as the "bricks". Meanwhile, an open-loop recycling liquid chromatography system will be built up for purification of target framework nucleic acids. And based on the purified high-order framework nucleic acids with uniform structure and integrated functions, an electrochemical platform will be developed for joint detection of multiple tumor markers of prostate cancer in pathological samples, so as to improve the diagnosis accuracy of prostatic cancer. Moreover, based on the results of joint detection and diagnosis, the clinical value of related tumor markers in the diagnosis of prostatic cancer will be clarified, so as to provide reliable detection principles for the early diagnosis, staging, therapeutic monitoring and postoperative evaluation of prostate cancer. The whole research work would provide new ideas and new technologies for the joint detection of multiple disease markers and the design, synthesis and purification of functional high-order framework nucleic acids, which would enrich the connotation of life analytical chemistry and related disciplines.

前列腺癌是发病率最高的癌症之一，实现前列腺癌的准确诊断成为降低前列腺癌死亡率、改善预后的关键。针对多基因参与、多信号转导通路共同作用的癌症诊断，相关多个标志物的联合检测比单一靶标分析准确性更高。结构精巧、功能可设计的框架核酸为生物界面的调控和高灵敏的生物分子传感提供了新手段。本项目拟先构建对单一前列腺肿瘤标志物特异性响应的功能化四面体框架核酸，并以此为“砖块”构建功能集成的高阶框架核酸结构；建立开路循环液相色谱体系及框架核酸的纯化方法；基于结构均一和功能集成的高阶框架核酸建立前列腺肿瘤多标志物联合电化学检测平台，进行实际病理样本的分析，提高前列腺癌诊断的准确性；明确相关标志物在前列腺癌诊断中临床价值，为前列腺癌早期诊断、分期、治疗监测及术后评价提供可靠的依据。项目的完成将为疾病多个标志物联合检测及功能化高阶框架核酸的设计合成与纯化提供新思路与新技术，丰富生命分析化学及相关学科的内涵。

前列腺癌是发病率最高的癌症之一，实现前列腺癌的准确诊断成为降低前列腺癌死亡率、改善预后的关键。针对多基因参与、多信号转导通路共同作用的癌症诊断，相关多个标志物的联合检测比单一靶标分析准确性更高。结构精巧、功能可设计的框架核酸为生物界面的调控和高灵敏的生物分子传感提供了新手段。因此，本项目提出构建对前列腺肿瘤标志物特异性响应的功能集成高阶框架核酸结构，建立前列腺癌准确检测方法，进行实际病理样本的分析，提高前列腺癌诊断的准确性。基于此，本项目以四面体框架核酸为基础单元构建了高阶框架核酸，实现了框架核酸数目可控组装及分子探针的细胞内高效运载，完成了前列腺癌细胞PC-3、MCF-7、A549等癌细胞和正常QSG-7701细胞中miRNA、ATP及凝血酶等多靶标联合检测与细胞成像分析。并基于血清样本中前列腺多个肿瘤标志物的联合检测实现了正常人、前列腺肿大及前列腺癌患者有效区分与诊断。同时针对疾病靶标检测灵敏度低等问题，设计构建了基于框架核酸及聚合酶链式反应的限域信号放大器及基于量子点的自驱动球形核酸信号放大器等，实现了前列腺及其他肿瘤细胞中相关标志物的超灵敏成像检测。相关研究成果为疾病多个标志物联合检测及功能化高阶框架核酸的设计合成提供了新思路与新技术，丰富了生命分析化学及相关学科的内涵。此外，依托于项目主体研究内容，项目基于金属有机框架的保护、装载能力，合成组装得到一系列AIE性质发射可调花状LMOFs及包裹量子点的有机金属框架纳米复合材料，并基于复合金属有机框架纳米材料构建了多个复杂体系多靶标传感检测平台，实现了生物及环境样本中疾病标志物及有害物质的快速灵敏检测，促进了分析化学、材料化学等相关学科的发展。项目实施期间总计在Anal. Chem., Chem. Commun.和Food Chem.等期刊上发表SCI论文11篇(均已标注资助)，培养博士生2名、硕士生2名。