【摘要】 目的以人肝癌细胞HepG2和SMMC-7721作为混合靶标筛选获得特异性核酸适配体并对其进行鉴定。方法以HepG2和SMMC-7721两种人源肝癌细胞为筛选混合靶标,利用细胞指数富集的配基系统进化(Cell-SELEX)技术筛选获得核酸适配体。利用流式细胞术检测文库富集情况,将富集文库进行克隆、测序,应用RNA structure、MEME在线软件进行序列分析,再次通过流式细胞术鉴定核酸适配体与靶标的结合能力及特异性。结果通过10轮Cell-SELEX筛选得到核酸适配体10-8,流式细胞术检测显示核酸适配体10-8与人源肝癌细胞系HepG2和SMMC-7721细胞结合,且特异性靶向肝癌细胞。结论筛选获得与人源肝癌细胞系HepG2和SMMC-7721双靶标特异性结合的核酸适配体10-8。更多还原

【Abstract】 Objective To perform the screening and identification of specific aptamers targeting both HepG2 and SMMC-7721 hepatoma cells. Methods Cell-SELEX was used to screen out the aptamers targeting both human HepG2 and SMMC-7721 hepatoma cells.Flow cytometry was used to evaluate the enrichment of pools,and the enriched pool was then cloned and sequenced.RNA structure and MEME online software were used for sequence analysis,and flow cytometry was used to identify the binding capacity and specificity of candidate aptamers to targets. Results The aptamer 10-8 was obtained after 10 rounds of Cell-SELEX.Flow cytometry showed that the aptamer 10-8 was bound to human HepG2 and SMMC-7721 hepatoma cell lines and specifically targeted hepatoma cells. Conclusion The aptamer 10-8 specifically binding to human HepG2 and SMMC-7721 hepatoma cells is obtained.更多还原