人骨肉瘤细胞特异性结合肽的筛选及验证  

Screening and identification of peptides specifically binding to human osteosarcoma cells

作　　者：卢坤[1] 姜勇[2] 管明强[1] 肖军[1] 王健[1] 李郅涵[1] 史占军[1] 

机构地区：[1]南方医科大学南方医院关节与骨病外科,广东广州510515 [2]南方医科大学病理生理教研室,广东广州510515

出　　处：《南方医科大学学报》2012年第5期647-650,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30371450)~~

摘　　要：目的获得人骨肉瘤细胞MG63特异性结合肽,为骨肉瘤靶向治疗奠定基础。方法应用噬菌体展示技术,以人骨肉瘤细胞作为靶细胞,293T细胞为差减细胞,筛选获得MG-63细胞特异性结合肽。酶联免疫法进行靶向性验证。应用荧光染色技术初步探讨短肽细胞受体位置。制作人骨肉瘤模型,尾静脉注射目标噬菌体,免疫组化检测其靶向性。结果经过四轮一步有机相离心分离方法,获得了与骨肉瘤细胞特异性结合的短肽,并测序获得其中出现次数最多的序列SLTNLSK,靶向验证结果显示该短肽有较强的特异性。结论应用噬菌体展示技术,获得了与人骨肉瘤细胞特异性结合的短肽,序列为SLTNLSK,并验证了其靶向性。可以作为骨肉瘤靶向治疗的导向性化合物。Objective To obtain the peptide that specifically binds to human osteosarcoma MG-63 cells from Ph.D.7TM phage display peptide library.Methods Human osteosarcoma MG-63 cells were used as the target cells with human embryonic kidney 293T cells as the control for screening the peptide from Ph.D.7TM phage display peptide library.The enriched specially binding peptides were verified by cell enzyme-linked immunosorbent assay（ELISA）.The location of the peptide in MG-63 cells was investigated using cell fluorescence staining,and targeting of the peptide was tested by organ immunohistochemistry with Osteosarcoma model.Results The specifically binding peptides were enriched after 4 rounds of screening.The sequence SLTNLSK was confirmed as the most frequent peptide by DNA sequencing and showed strong specificity verified by cell ELISA,fluorescent staining and organ immunohistochemistry.Conclusion A peptide that specifically binds to MG-63 cells has been screened from Ph.D.7TM phage display peptide library to serve as a potential candidate for osteosarcoma-targeting therapy.

关 键 词：骨肉瘤 靶向治疗 噬菌体展示技术 结合肽 

分 类 号：R738[医药卫生—肿瘤]