P-糖蛋白结合肽的筛选及合成肽鉴定

Screening and synthesis of specific peptide to P-glycoprotein

目的筛选人膀胱癌P-糖蛋白特异性结合肽并合成鉴定。方法利用表达获得的P-糖蛋白胞外段融合蛋白为靶蛋白,采用酶联板法筛选噬菌体随机12肽库,化学合成该特异性肽序列,免疫细胞化学方法进行鉴定。结果从噬菌体随机肽库中筛选获得了与P-糖蛋白特异性结合的噬菌体阳性克隆,测序获得特异性结合肽序列。免疫细胞化学结果显示特异性合成肽可与耐药细胞BIU-87/ADM结合,而与敏感细胞BIU-87不结合。结论筛选获得的结合肽具有一定的亲和力,可与特定的肿瘤细胞结合,表现出一定的肿瘤特异性;P-糖蛋白结合肽的筛选,为人膀胱癌多药耐药的靶向治疗结论提供实验依据。

Objective To screen and identify peptide specific to P-glycoprotein. Methods The fused P-glycoprotein was used as target to biospan its binding peptide with enzyme-linked plate assay in phage random 12 peptide library,synthesis of peptide specific to P-glycoprotein and immunocytochemical staining in BIU-87/ADM and BIU-87 cells was done to identify the posive phage clones. Results Peptide specific binding to P-glycoprotein was acquired after biospanning. The inserted sequence of the peptide was identified by sequencing analyses. Immunocytochemical staining results revealed that the binding only occurred in MDR cells BIU-87/ADM,not in sensitive cells BIU-87. Conclusion It indicates that this kind of binding exerts its affinity and specificity to MDR cells. The biospanning of peptide provides a base for the targeted reversal of multidrug resistance in human bladder cancer.