转移性大肠癌特异性核酸适配体分子信标的构建及其靶向成像  

Construction of specific molecular aptamer beacon for metastatic colorectal cancer and its targeting imaging

作　　者：鲁妍兵 李婉明[1] 方瑾[1] LU Yan-bing;LI Wan-ming;FANG Jin(Department of Molecular Cell Biology,Key Laboratory of Cell Biology,Ministry of Public Health,and Key Laboratory of Medical Cell Biology,Ministry of Education,China Medical University,Shenyang 110122,China)

机构地区：[1]中国医科大学卫生部细胞生物学重点实验室,医学细胞生物学教育部重点实验室,生命科学学院分子细胞生物学教研室,辽宁沈阳110122

出　　处：《解剖科学进展》2022年第5期523-526,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81672920,81502703);沈阳市中青年科技创新人才支持计划(BC190261)。

摘　　要：目的设计并构建靶向转移性大肠癌的核酸适配体分子信标(MAB),分析其生物活性并进行特异性成像研究,为转移性大肠癌的靶向成像提供新的方法。方法根据分子信标的设计原则在转移性大肠癌特异性核酸适配体的基础上构建MAB,采用流式细胞术对MAB的特异性和亲和性进行分析,并对不同温度下(4℃、25℃和37℃)MAB与靶细胞的结合情况进行检测;采用激光共聚焦显微镜对MAB与靶细胞靶向成像进行观察。结果构建的MAB与转移性大肠癌细胞的结合具有高亲和性和高特异性,解离常数为(22.70±2.46)nM,且不受温度的影响,激光共聚焦显微镜观察结果表明MAB不需洗涤过程便可一步法实现转移性大肠癌细胞的靶向成像。结论成功构建出基于转移性大肠癌特异性核酸适配体的MAB,建立了一种快速、便捷的转移性大肠癌的靶向成像方法。Objective Molecular aptamer beacon targeting metastatic colorectal cancer was designed and constructed,and its biological activity was analyzed and specific imaging study was carried out,providing a new method for targeted imaging of metastatic colorectal cancer.Methods According to the design principle of molecular beacon,the molecular aptamer beacon was constructed on the aptamer targeting metastatic colorectal cancer,the specificity and the affinity of MAB were analyzed by flow cytometry,and the binding of MAB to target cells was detected by flow cytometry at different temperatures(4℃,25℃ and 37℃),the targeted imaging of MAB and target cells was observed by laser confocal microscope.Results The binding of the constructed MAB has high affinity and specificity with metastatic colorectal cancer cells,and the dissociation constant was(22.70±2.46)nM,and was not affected by temperature.MAB could achieve targeted imaging of metastatic colorectal cancer cells in one step without washing.Conclusion The MAB,based on aptamer targeting metastatic colorectal cancer,was constructed successfully,a rapid and convenient target imaging method for metastatic colorectal cancer was established.

关 键 词：核酸适配体 分子信标 大肠癌 靶向成像 

分 类 号：R735.34[医药卫生—肿瘤]