无酶级联催化发夹自组装和杂交链反应信号放大细胞表面特定蛋白糖基化高灵敏成像分析和高效光动力治疗  

Visualization of Protein-specific Glycosylation and Photodynamic Therapy via an Enzyme-free Amplification Strategy of Catalytic Hairpin Assembly and Hybridization Chain Reaction

作　　者：李婷[1] 邢思敏 刘洋[1] LI Ting;XING Simin;LIU Yang(Department of Chemistry,Beijing Key Laboratory for Analytical Methods and Instrumentation,Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology,Ministry of Education,Tsinghua University,Beijing 100084,China)

机构地区：[1]清华大学化学系,生命有机磷化学及化学生物学教育部重点实验室,微量分析测试方法与仪器研制北京市重点实验室,北京100084

出　　处：《高等学校化学学报》2023年第8期54-65,共12页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:22174084)资助.

摘　　要：开发了一种基于邻近诱导的催化发夹组装和杂交链反应(CHA-HCR)的高灵敏度、无酶信号放大策略,用于癌症细胞原位蛋白质特异性糖基化高灵敏荧光成像和高效光动力治疗.设计了2种DNA探针,分别用于靶蛋白和聚糖的特异性标记,其中蛋白探针(PP)通过适配体识别MUC1蛋白,聚糖探针(GP)通过生物正交反应特异性结合唾液酸.由于邻近效应,PP中的CHA起始序列与相邻的GP杂交,并引发随后的CHA-HCR级联反应.最后,在靶蛋白的特定糖基化上形成具有多个荧光分子或光敏剂的延伸双链DNA,实现了细胞表面特定糖蛋白的高灵敏选择性成像分析和肿瘤细胞灭杀,为癌症的诊断和治疗提供了一条通用途径,也为通过靶向特定糖蛋白增强细胞毒效应的光动力治疗提供了工具.该策略为揭示糖基化机制、筛选聚糖相关生物标志物和开发靶向疗法提供了参考.Herein,a highly sensitive,enzyme-free signal amplification strategy based on the proximity-induced catalytic hairpin assembly and hybridization chain reaction(CHA-HCR)was developed for protein-specific glycosylation fluorescence imaging of cancer cells in situ and highly efficient photodynamic therapy.Two DNA probes were designed for the specific labeling of target proteins and glycans,respectively,where the protein probe(PP)identified the MUC1 protein by aptamer and the glycan probe(GP)specifically bound to sialic acid via bioorthogonal reaction.Due to the proximity effect,the CHA initiation sequence in the PP hybridized with the adjacent GPs and triggered a subsequent CHA-HCR cascade reaction.An extended double-stranded DNA strand with multiple fluorescent molecules or photosensitizers formed on specific glycosylation of the target protein,which provided a universal avenue for the multiple glycosylation imaging on the cancer cell surface with high sensitivity and a valuable way for photodynamic therapy with enhanced cytotoxic effect by targeting the specific glycoproteins.This strategy supplies the reference for uncovering the mechanism of glycosylation,screening glycan-related biomarkers and developing targeted therapies.

关 键 词：邻近 生物正交反应 无酶信号放大 糖基化成像 光动力治疗 

分 类 号：O652[理学—分析化学]