血清对细胞摄取DNA四面体纳米结构行为的影响

摘要

研究了细胞培养基中的胎牛血清(FBS)对DNA四面体(Tetrahedral DNA nanostructure,TDNs)进入HeLa细胞的速度和内吞途径的影响.采用自组装技术得到荧光标记的TDNs结构,利用HPLC技术分离得到纯度>95%的TDNs单体,分别采用流式细胞术和共聚焦显微成像等技术比较了在有无血清的情况下,细胞摄取量随时间的变化以及FBS对TDNs摄取途径的影响.实验结果表明,TDNs在培养基和细胞裂解液环境中可以稳定存在12 h以上,培养基中的FBS能够提高HeLa细胞对四面体的摄取量, 但并未改变TDNs进入HeLa细胞的内吞途径.本研究揭示了环境中蛋白质等生物分子对于DNA四面体结构与细胞界面相互作用的影响,为基于DNA纳米材料的细胞学纳米载体的设计和优化提供了新思路.%In the present study, the effect of serum (fetal bovine serum, FBS) on cellular uptake behaviors of DNA tetrahedron (TDNs) was investigated.Fluorescence-labeled TDNs were synthesized by self-assembly and purified with HPLC to achieve TDNs product with purity of 95%.By means of flow cytometry and confocal microscopy, the kinetics of TDNs endocytosis in HeLa cells in the presence or absence of FBS was compared.The results showed that TDNs remained intact in complete medium and cell lysate for more than 12 hours.FBS increased the amount of internalized TDNs in HeLa cells, whereas it did not change the route of caveolin-dependent endocytosis.Herein, our study provided a new insight into the effects of biomolecules on the interaction between cells and DNA nanostructures, which helped the future development of DNA-based intracellular nanocarriers.