P-糖蛋白的核酸适配体筛选及应用研究  

Research on Selecting of P-gp Aptamers and Its Application

作　　者：李玉娟[1] 梁敏 武广霞 孙欣欣[1] 刘珊 邓玉林[1] LI Yujuan;LIANG Min;WU Guangxia;SUN Xinxin;LIU Shan;DENG Yulin(School of Life Science,Beijing Institute of Technology,Beijing 100081,China)

机构地区：[1]北京理工大学生命学院,北京100081

出　　处：《北京理工大学学报》2021年第11期1236-1244,共9页Transactions of Beijing Institute of Technology

基　　金：国家自然科学基金资助项目(81573693,81973572)。

摘　　要：基于SELEX技术筛选P-糖蛋白(permeability glycoprotein,P-gp)的特异性核酸适配体(aptamer,Apt),并用酶联免疫吸附测定(ELISA)法、表面等离子体共振(SPR)法表征了Apt与P-gp的结合能力,采用与P-gp结合能力最强的Apt5在人结直肠腺癌细胞(Caco-2)、人脑微管内皮细胞(hCMEC/D3)两种细胞系上考察了对P-gp外排功能的抑制效果.筛选得到10条长度为81 bp的P-gp核酸适配体,结合能力大小顺序为:Apt5>Apt8>Apt1>Apt4>Apt6>Apt10>Apt7>Apt2>Apt9>Apt3.Apt5使hCMEC/D3及Caco-2细胞内罗丹明123的蓄积量分别增加了40.77%(P<0.01),32.39%(P<0.05),能潜在抑制P-gp的外排功能.本研究首次报道了P-gp的多条核酸适配体,与P-gp有较强的结合能力,并能抑制P-gp的外排功能,有望成为P-gp的潜在新型抑制剂.The SELEX technology was used to select aptamers(Apt)of permeability glycoprotein(P-gp).The enzyme linked immunosorbent assay(ELISA)and surface plasmon resonance(SPR)methods were used to determine the affinity between Apts and P-gp.Apt5 was chosen to evaluate the inhibitory effect on P-gp efflux function with immortalized human brain capillary endothelial cells(hCMEC/D3)and human epithelial colorectal adenocarcinoma cells(Caco-2).As a result,ten of 81bp Apts were selected.The affinity order of Apt was Apt5>Apt8>Apt1>Apt4>Apt6>Apt10>Apt7>Apt2>Apt9>Apt3.Further research showed that Apt5 significantly increased the cellular accumulation of rhodamine 123 both in hCMEC/D3 and Caco-2 cells,with a fold of 40.77%(P<0.01)and 32.39%(P<0.05),respectively,which indicated that Apt5potentially inhibited the efflux function of P-gp.In present study,several Apt of P-gp are presented,which had high affinity with P-gp and could inhibit P-gp efflux function.These Apts might be potential novel P-gp inhibitor.

关 键 词：P-糖蛋白 SELEX 核酸适配体 P-糖蛋白外排功能 抑制剂 

分 类 号：R963[医药卫生—微生物与生化药学]