噬菌体随机肽库中与神经胶质瘤细胞系SWO-38特异性靶向结合短肽的筛选  

Selection of Targeted Glioblastoma Tumor Cell - bindng and Internalizing Peptides through Phage Display Vector

作　　者：王冰[1] 钟雪云[1] 秦艳芳[1] 钟瑛[1] 

机构地区：[1]广州暨南大学医学院病理教研室,广东广州510632

出　　处：《中国神经肿瘤杂志》2004年第1期26-31,共6页Chinese Journal of Neuro-Oncology

基　　金：国家科技部重大基础研究前期研究专项(2002ccc0400);广州市科技局科研重点立项课题(2001-E-01-2)

摘　　要：神经胶质瘤的常规治疗尚难取得令人满意的效果,寻找高效和高特异性杀伤肿瘤细胞的药物已成为提高其治愈率的研究热点。本研究旨在利用噬菌体随机肽库找到与神经胶质瘤细胞系SWO-38特异性结合并内化人细胞的短肽序列。方法:利用噬菌体随机12肽库对肿瘤细胞进行5轮全细胞筛选,分析筛选后单克隆对神经胶质瘤细胞的特异性结合能力并进行免疫荧光分析。提取单克隆DNA,测序,得出短肽序列进行比对分析。结果:通过5轮筛选后的噬菌体库及所挑选的表面及内化部分各13个单克隆均显示对胶质瘤细胞有较高的特异性结合,并测序得到三条重复性高的多肽序列。免疫荧光分析显示噬菌体大量聚集在细胞表面并能够内化进入到细胞内。结论:通过噬菌体随机肽库对肿瘤细胞进行全细胞筛选得到的噬菌体多肽能高特异性与胶质瘤瘤细胞SWO-38结合,可作为肿瘤导向药物研究的载体。BACKGROUND & OBJECTIVE: It has been investigated for a long time for highly effective and specific agents to cure gliomas. This study was to isolate peptides that showed highly selective binding and internaliz-ing into glioblastoma cell line SWO-38. METHODS: The tumor cells were screened by five rounds of whole cell screen through the Ph. D.-12 phage display library. The monoclones specific binding efficiency to the tumor cell was analyzed. The DNA of phages was extracted, sequenced and translated into amino acids. Through immunofluorescence microscopy, we selected the phage particles which bound on cell membrane and internalized into cytoplast. RESULTS: After five rounds of screen, all of the monoclones showed highly specific binding to SWO-38 cells. Through sequenc-ing, we found three repeated peptide sequences. CONCLUSION: Whole cell screening in tumor cells through random phage peptide library can obtain phage peptides with highly specific binding and internalizing ability. The peptides could used as therapeutic vector for targeted drug deliv-ery.

关 键 词：噬菌体肽库 神经胶质瘤 细胞系 SWO-38 短肽 肿瘤细胞 肿瘤导向 

分 类 号：R739.41[医药卫生—肿瘤]