与卵巢癌细胞表面受体特异性结合的多肽片段研究  

Selection of targets locating on the surface of epithelial ovarian cancer cells by using phage peptide library

作　　者：王世宣[1] 周洪源[1] 马湘一[1] 陈刚[1] 卢运萍[1] 何福仙[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《中华妇产科杂志》2003年第7期412-414,共3页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目 (3 0 170 976);973工程基金资助项目 (2 0 0 2CB5 13 10 0 )

摘　　要：目的　从噬菌体随机肽库中 ,筛选与卵巢癌细胞特异性结合的多肽片段 ,为卵巢癌靶向治疗提供实验基础。方法　从噬菌体随机肽库中 ,依次筛选 4株卵巢癌细胞株及正常卵巢细胞 ,并获得阳性克隆 ,再经过酶联免疫吸附实验挑选出目的克隆 ,进行DNA测序及同源性分析。结果　将获得的 10个目的克隆 ,进行DNA序列分析 ,得到 1段可以与卵巢癌细胞结合的特异性多肽片段(YYGLAEVDAGGS)。结论　噬菌体随机肽库为卵巢癌细胞表面靶位的筛选提供了高效、快捷的筛选体系 ,实验所获得的特异性多肽片段 。Objective The aim of this study was to select specific targets locating on the surface of epithelial ovarian cancer cells Methods Peptide phage display library was used to isolate specific ligand to ovarian cancer cell receptors.The diluted library was incubated with the normal ovarian cells that primarily cultured before hand, and then the supernatant of nonbonding phage was added to the first epithelial ovarian cancer cell line A2780.Phage that binds to the cell surface are eluted and then amplified to be used in the next round. After the third round of panning,the elute was used as input phage for the next cell line biopanning. Four epithelial ovarian cancer cell lines were used one by one in this way. Finally the positive clones were identified by ELISA assay. Sequencing analysis was carried out for further identification Results 10 positive clones were chosen and one was regarded as target clone. A candidate sequence (YYGLAEVDAGGS)was identified by amino acid sequence assay Conclusion The phage peptide library provides an efficient selection system for searching special targets locating on the cell surface.

关 键 词：卵巢肿瘤 肽库 肽类 细胞表面受体 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]