细胞表面EpCAM糖基化程度差异是导致健康与恶性组织中细胞分子功能差异的重要因素，与乳腺癌、肝癌、结肠癌等多种癌症的迁移侵袭和预后密切相关。然而，当前针对EpCAM低糖基化糖型的原位检测仍十分困难，且细胞分子功能机制尚不清楚。本项目拟基于适配体介导的局域动态DNA纳米自组装技术，设计并构建高特异性的“DNA分级编码”和“DNA纳米马达”蛋白糖型信号甄别策略；再结合滚环扩增和交换反应信号扩增的DNA信号放大技术，建立高灵敏的蛋白糖型信号收集系统。发展原创性EpCAM低糖基化糖型原位、高效、实时、多通道的检测新方法。进一步探索方法对肿瘤迁移侵袭过程中EpCAM糖型的动态监测，并用于临床乳腺癌组织样本EpCAM糖基化水平的评估。本项目的实施将为探究EpCAM在肿瘤发生发展过程中的作用、筛选新型肿瘤标志物及甄定治疗靶点提供有力工具，为乳腺癌等肿瘤的精确分子分型、精准诊疗和预后判断提供全新技术平台。

The differential glycosylation of EpCAM could be an important factor that brings about differences in the function of EpCAM in healthy versus malignant tissue. EpCAM glycosylation has been found to be reliably associated with greater migration, invasion and prognosis in some tumors including breast cancer, liver cancer, colon cancer, etc. However, the in situ monitoring of EpCAM-bound hypoglycosylated glycoforms is still quite difficult, and the functional mechanism of cell molecules remains unclear. This project intends to design and construct highly specific “DNA hierarchical coding” and “DNA nanomotor” protein-bound glycoform signal screening strategies based on aptamer-mediated localized dynamic DNA nanoassembly technology. Combined with rolling circle amplification and signal amplification by exchange reaction, highly sensitive protein-bound glycoform signal collection systems are established. Original methods are proposed for in situ, efficient, real-time, multi-channel detection of EpCAM-bound hypoglycosylated glycoforms. The developed protocols are employed for dynamic monitoring of the glycoform variations during tumor migration and invasion, further explored to the evaluation of EpCAM glycosylation levels in clinical breast cancer tissues. This project will not only provide a powerful tool for exploring the role of EpCAM in the development of tumors, screening new tumor markers and identifying therapeutic targets, but a novel platform for accurate molecular subtyping, precise diagnosis and prognosis of breast cancer and other tumors.