噬菌体展示肽库筛选大肠癌细胞特异性结合肽的实验研究  

Screening peptides binding specifically to large intestinal cancer LoVo cells from phage random peptide library

作　　者：廖康雄[1] 姚学清[2] 吴承堂[1] 林锋[2] 吴伍林[2] 曾穗德[2] 罗育其[1] 雷尚通[1] 

机构地区：[1]南方医科大学南方医院普通外科,广东广州510515 [2]广东省人民医院胃肠外科,广东广州510080

出　　处：《南方医科大学学报》2008年第6期986-990,共5页Journal of Southern Medical University

基　　金：广东省自然科学基金博士启动项目(5301056)~~

摘　　要：目的筛选人源大肠癌LoVo细胞株特异结合的短肽,作为大肠癌靶向治疗的载体。方法以大肠癌LoVo细胞株作为靶细胞,人正常结肠黏膜上皮细胞为吸附细胞对噬菌体展示环七肽库进行差减筛选,采用细胞ELISA与免疫荧光检测鉴定噬菌体阳性克隆并测序。结果对噬菌体展示环七肽库进行3轮筛选后,从随机挑取的20个噬菌体克隆中获得5个能与大肠癌LoVo细胞特异结合,而不与人正常结肠黏膜上皮细胞结合的阳性克隆,其氨基酸序列含有保守序列RPMP。结论利用噬菌体展示肽库技术,可以成功筛选到大肠癌细胞的特异性结合肽,可能成为大肠癌靶向治疗的载体。Objective To screen the polypeptides specifically binding to human large intestinal cancer.LoVo cells from a phage-displayed peptide library for potential use as targeting vectors for large intestinal cancer therapy, Methods With the LoVo cells as the target cells and human normal large intestinal mucosal epithelial cells as the absorber cells for subtraction biopanning from a c7c phage-display peptide library, the positive phage clones were identified by enzyme-linked immunosorbent assay （ELISA） and immunofluorescence detection. The amino acid sequences of the identified peptides were deduced by DNA sequencing. Results After 3 rounds of screening, 5 positive phage clones showing specific binding to LoVo cells and containing conserved motif RPMP were obtained from the 20 randomly selected clones. Conclusion Specific peptide against large intestinal cancer cells can be obtained from a phage-display peptide library for use as potential vectors for targeting therapy of large intestinal cancer.

关 键 词：噬菌体肽库 LOVO细胞 差减筛选 大肠癌 M13 

分 类 号：R735.34[医药卫生—肿瘤]