基于DNA自组装信号放大策略的肝细胞癌肿瘤标志物电化学检测方法研究

Hepatocellular carcinoma (HCC) is a common type of digestive tract cancer with high morbidity and mortality. A hope to decrease deaths from HCC is offered by earliest possible diagnose. Recent advances in molecular biology elucidate that increased levels of tumor markers in human serum are associated in patients with certain tumors. Therefore, the sensitive and accurate assays for tumor markers will be of great value for cancer diagnosis. Up to now, many efforts have been made for the detection of HCC tumor markers. However, shortcomings with the sensitivity and anti-interference ability of existing methods limit their application in early HCC diagnosis. DNA self-assembly based signal amplification is a newly developed enzyme-free technique that has proven useful in significantly amplifying signals. By using antibodies or aptamers as the recognition elements, this project will develop novel electrochemical methods with ultra-highly clinical sensitivity and selectivity for detecting HCC tumor markers via DNA self-assembly based signal amplification, providing technical support for early diagnosis of HCC.

肝细胞癌是一种具有高发病率和高致死率的消化道恶性肿瘤。尽早诊断是降低肝细胞癌死亡率的关键。肿瘤标志物是肿瘤组织异样表达的一类化学物质，其在血清中的存在或量变可以提示肿瘤的发生，是肿瘤早期诊断的重要指标。然而，现有的肝细胞癌肿瘤标志物检测方法普遍具有灵敏度不够高或抗干扰能力差等缺陷，难以满足早期诊断的需要。另一方面，DNA自组装信号放大策略作为一种恒温、无酶的新兴信号放大技术，在生物分析中表现出广阔的应用前景。在此背景下，本项目拟以电化学技术为检测手段，以抗体或核酸适体为识别分子，将DNA自组装信号放大策略与蛋白质检测相结合，发展能够满足血清学诊断需求的高度灵敏和特异性的肝细胞癌肿瘤标志物检测新方法，以期为肝细胞癌的早期诊断提供可能的帮助。

肝细胞癌是最常见的消化道恶性肿瘤之一，也是人类第三大癌症死亡原因。近年来，肿瘤标志物的灵敏检测被认为是实现肝细胞癌诊断和预后，进而提高肝细胞癌患者存活率的重要手段。在本项目中，我们借助于一种恒温、无酶的新兴信号放大技术，即DNA自组装信号放大技术，选择若干种具有肝细胞癌诊断价值的肿瘤标志物作为研究对象，开展高灵敏的电化学检测新方法研究。结合分子识别、点击化学以及电化学传感等领域的最新研究成果，本项目通过长距链式自组装、级联链置换反应、催化发卡环组装等DNA自组装信号放大策略的应用，设计了多种适用性强且效果显著的电化学检测方案，并由此实现了以干扰素-γ、磷脂酰肌醇蛋白聚糖3、细胞表面生物素受体、肝细胞癌相关基因MXR7等为代表的肝细胞癌肿瘤标志物的高灵敏检测。在国家自然基金的资助下，相关研究工作开展顺利，取得了较好的成果。三年里，一共发表SCI论文7篇（均为第一作者或通讯作者；其中，影响因子大于5的3篇）；申请和授权专利2项；协助培养硕士研究生7名（其中已毕业3名）；顺利实现了申请时的预期目标。