基于Cell-SELEX技术对人B细胞淋巴瘤SU-DHL细胞适配体的筛选与构建  

Screening and Construction of SU-DHL Aptamers for Human B-cell Lymphoma Based on Cell-SELEX Technology

作　　者：于文莹 王梦主 杜冰洋[3] 王昶[3] 高丽君 夏薇 YU Wenying;WANG Mengzhu;DU Bingyang;WANG Chang;GAO Lijun;XIA Wei(College of Medical Technology,Beihua University,Jilin 132013,China;Xiantao First People’s Hospital,Yangtze University,Xiantao 433000,China;Jilin Central Hospital,Jilin 132011,China)

机构地区：[1]北华大学医学技术学院,吉林吉林132013 [2]长江大学附属仙桃市第一人民医院,湖北仙桃433000 [3]吉林市中心医院,吉林吉林132011

出　　处：《北华大学学报（自然科学版）》2023年第4期449-453,共5页Journal of Beihua University（Natural Science）

基　　金：吉林省科技厅重点科技攻关项目(20140204016YY);吉林市科技发展计划项目民生科技创新专项(2015334009).

摘　　要：目的筛选与淋巴瘤SU-DHL-8细胞特异性结合的适配体,用于淋巴瘤的早期诊断.方法利用76 bp的随机单链DNA文库,以SU-DHL-8细胞为靶细胞、Ramos细胞为消减细胞,采用以细胞为靶标的指数富集配体系统进化(cell based systematic evolution of ligands exponential enrichment,Cell-SELEX)技术筛选富集与靶细胞结合的适配体;利用PCR及生物素-链霉亲和素层析技术制备次级文库,经多轮筛选,最终筛选出与SU-DHL-8细胞特异性结合的适配体.结果经过11轮正向筛选及2轮反向筛选,次级ssDNA文库与SU-DHL-8细胞的亲和力逐渐增高,第12轮和第13轮的荧光曲线基本重合,说明SU-DHL-8细胞的特异适配体构建成功.结论应用Cell-SELEX技术可成功筛选出淋巴瘤SU-DHL-8细胞的特异适配体,为淋巴瘤的精确诊断和早期诊断提供依据.Objective To screen the aptamers that specifically bind to lymphoma SU-DHL-8 cells for early diagnosis of lymphoma.Method 76 bp random single-stranded DNA library was used,with SU-DHL-8 cells as the target cells and Ramos cells as the subtractive cells.The ligand system evolution technology with exponential enrichment was used to screen and enrich the aptamers that bind to the target cells;The secondary library was prepared by PCR and biotin-streptavidin chromatography.After multiple rounds of screening,the aptamer specifically binding to SU-DHL-8 cells was finally selected.Results After 11 rounds of forward screening and 2 rounds of reverse screening,the affinity between the secondary ssDNA library and SU-DHL-8 cells gradually increased,and the fluorescence curves of the 12th and 13th rounds basically coincided,indicating that the specific aptamer of SU-DHL-8 cells was successfully constructed.Conclusion The Cell-SELEX technique can successfully screen the specific aptamer of lymphoma SU-DHL-8 cells,and the new diagnostic marker of lymphoma can be obtained by labeling the probe,which provides the basis for accurate and early diagnosis of lymphoma.

关 键 词：SU-DHL-8细胞 淋巴瘤 适配体 SELEX技术 

分 类 号：R735.3[医药卫生—肿瘤]