乳腺癌进展期相关细胞糖基化原位电化学监测

The glycosylation of tumor cells is closely related to cell physiological state and the development of disease. However, the major factor that restricts the study of glycosylation is the lack of effective detection methods. This project will use the green, stable nanometer material surface modification and biological functionalization method, combined with sensitive and fast electrochemical detection technique and flow cytometry, to develop a series of low cost, sensitive and high specific methods for recognizing and dynamically monitoring of breast cancer cell surface glycans, and further dynamically monitoring the glycosylation of breast cancer cells at different progressive stage, in order to reveal the relationship between progressive stage and glycosylation of breast cancer cells. The obtained in situ method based on nano-sized mimic recognization interface will be important for revealing the influence of anti-cancer drug and carcinogen on cell surface glycan expression, investigating the interaction of drug (or carcinogen) and glycan (or glycoprotein), studying the function of glycan and its receptor in cell communication, cell signal transduction, apoptosis, canceration, tumorigenesis, tumor development and transfer. This project will establish the foundation for the early diagnosis of breast cancer based on the detection of cell surface glycans, and provide a possible avenue for the cure of tumor diseases.

肿瘤细胞表面的糖基化模式与细胞的生理状况和疾病的发生发展密切相关。目前制约糖基化研究的主要因素是缺少有效的分析检测方法。申请者近几年将电化学分析技术应用于肿瘤细胞表面聚糖的原位定量检测取得一定进展。本项目拟通过绿色、稳定的纳米材料表面修饰和生物功能化方法，高效、高选择性的糖基识别策略，结合灵敏、快速的电化学检测技术和流式细胞技术，建立一系列全新、价廉、高灵敏度、高特异性的乳腺癌细胞表面糖基识别、检测和动态监测新方法；通过引入标准化荧光微珠，建立细胞表面聚糖定量流式细胞检测方法，与电化学方法进行验证；对临床不同进展期乳腺癌标本进行分析，实现不同进展期乳腺癌细胞糖基化的动态监测，以揭示乳腺癌发展程度与癌细胞糖基化水平的密切联系；进一步研究抗癌药物对细胞膜上糖基表达的影响，探讨药物与糖基及糖基化蛋白的相互作用。本项目的研究将为发展基于细胞表面糖基检测的乳腺癌早期诊断新技术奠定基础。

肿瘤细胞异常的糖基化与癌症发生进展密切相关。本项目成功利用凝集素对多壁纳米管进行表面修饰，得到对糖基具有高选择性的识别能力的纳米复合物，结合灵敏、快速的电化学定量检测技术，建立了价廉、高灵敏度、高特异性的乳腺癌细胞表面糖基电化学方法检测；进一步结合双通道丝网印刷电极，构建了双通道的糖基检测电化学阵列。同时通过构建基于凝集素的对糖基特异性是别的量子点纳米探针，结合量子点优良的光学性质，建立了原位均相检测乳腺癌细胞膜糖基的荧光定量检测方法。光谱学、形态学、电化学、细胞学等一系列实验证实了所建立的方法具备宽的检测范围，低的检测限，良好的重复性和稳定性，操作简便快捷，完全适合应用于临床组织样本的检测。进一步将所建立的糖基检测方法应用于临床样本分析，实现了不同进展期乳腺癌细胞糖基化的动态监测，揭示了乳腺癌发展程度与癌细胞糖基化水平的联系，提出了可能的乳腺癌进展期相关的生物标志物，为临床乳腺癌的防治提供了新的工具。