基于长余辉发光纳米探针的前列腺癌细胞表面聚糖原位分析

Cell surface glycans play an essential role in various cellular processes including molecular recognition, cell communication, immunoresponse, cell adhesion and migration.It has been demonstrated that aberration of surface glycans is closely related to a lot of fatal diseases such as cancers. In recent years, cancer glycobiology has attracted more and more attentions of worldwide researchers.Since the inherent complexity of glycans, there still remains a great challenge for detection and analysis of cell surface glycans, especially in situ analysis on living cells. Based on the applicant's research experience, this project will innovatively develop a kind of persistent luminescence nanoparticles-based optical probe for in situ, quantitative, highly sensitive, and excitation-free analysis of cancer cell surface glycans. In this project,normal and prostate cancer cell lines are selected as research model. The following subprojects will be investigated in detail: microfluidic lab on a chip is employed to profile glycans on the surface of normal prostate cells and prostate cancer cells; study anti-cancer drugs-induced surface glycan expression and structures changes and further investigate the effect of glycans' aberration on cancer cell proliferation,adhesion,invasion and metastasis; the glycan changes on specific membrane glycoproteins are investigated with antibody microarrays to understand the role of surface glycans in prostate cancer progression and its molecular mechnism.In a word,this project not only provides a novel and powerful technique for in situ imaging and analysis of cell surface glycans, accelerates the screening of cancer biomarkers and early cancer diagnosis, but also renders useful fundemental information for anti-cancer drug discovery and targetted cancer therapy.

细胞表面聚糖在细胞生命活动中具有极其重要的作用，研究表明聚糖异常与很多重大疾病如肿瘤密切相关，近年来肿瘤糖生物学日益成为国内外研究的热点。由于聚糖本身的高度复杂性，聚糖的分析检测，尤其是原位成像与解析进展缓慢。申请人在前期研究基础上，创新性地构建一种长余辉发光纳米探针，实现细胞表面聚糖的原位、定量、高灵敏、免激发分析。本项目以不同类型的前列腺癌细胞系为模型，开展以下研究：应用微流控芯片分析正常细胞和不同阶段肿瘤细胞表面聚糖表达和结构的差异；研究抗肿瘤药物对细胞表面糖基表达谱的影响，探讨表面聚糖改变与肿瘤细胞增殖、粘附和迁移的关系；应用抗体微阵列芯片研究前列腺癌细胞表面特定膜糖蛋白的糖基化，从分子水平上了解聚糖异常与前列腺癌的关系及其作用机制。本项目不仅可以为肿瘤细胞表面聚糖的检测和解析提供新的解决途径，促进肿瘤标志物的筛选与肿瘤早期诊断，还可为肿瘤的靶向治疗和药物开发提供理论依据。

细胞表面糖链是细胞与外界沟通的重要信号分子，在细胞生命活动中起着十分重要的作用。已有研究表明，细胞表面糖链的异常与肿瘤的发生、发展密切相关。本项目旨在开发一种长余辉发光纳米材料，并将其与特异性的糖基识别分子相结合，构建新型光学纳米探针，应用于细胞内糖蛋白和表面糖基的高灵敏、免激发成像及分析，进而探讨细胞糖基化异常（糖基化水平和糖基种类等）与细胞的生物学功能的关系。在本项目的资助下，项目负责人及其团队取得了如下成果：成功合成了ZGC长余辉纳米材料，并进行了系统表征，确认了材料的形貌、晶体结构、光学特性等理化性质；首次实现了液相化学掺杂法对其光学性质进行调控，并初步掌握了相关机理；基于所合成的长余辉纳米探针成功构建了生物分子标记和非标记检测分析策略，并实现了高灵敏免疫分析和正常细胞/肿瘤细胞表面聚糖表达谱的定性分析；评价了长余辉材料ZGC的血液相容性，并通过亲水性聚合物聚乙二醇（PEG）修饰，有效提高了ZGC的血液相容性，为该材料在体内生物成像和疾病治疗提供了理论基础和切实可行的解决方法；应用微阵列免疫芯片和微流控生物芯片开展了蛋白糖基化修饰的高灵敏分析，并进一步探究了O-GlcNAc糖基化水平改变对细胞增殖和迁移的影响。本课题成功开发了基于长余辉纳米探针的细胞表面糖链原位检测和分析方法，对认识糖基化异常与肿瘤发生发展的关系及肿瘤的早期检测具有重要意义；另外，本课题开发了一种简单有效的长余辉材料光学性质调控策略，并评价和改善了材料的血液相容性，为拓展长余辉纳米材料的体内外应用奠定了基础。