肝癌特异性噬菌体多肽的筛选和初步鉴定  

Screening and preliminary identification of liver cancer specific peptide from a phage display peptide library

作　　者：罗时敏[1] 臧林泉[2] 

机构地区：[1]广州市第一人民医院综合外科,510180 [2]广东药学院药科学院药理教研室,广州510006

出　　处：《中华肝胆外科杂志》2009年第12期928-930,共3页Chinese Journal of Hepatobiliary Surgery

基　　金：本课题受国家自然科学基金（30572177）和教育部重点基金（208105）资助

摘　　要：目的利用噬菌体展示肽库筛选与肝癌HepG2细胞特异性结合的多肽，为筛选及明确新的肝癌早期诊断和治疗标志物打下基础。方法以肝癌细胞HepG2为靶细胞，LO-2为吸附细胞，在37℃条件下对噬菌体随机12肽库进行多轮减性筛选，挑取单克隆扩增并鉴定。利用ELISA初步鉴定克隆亲和力，测定阳性克隆DNA测序并进行同源性及氨基酸分析。结果经过3轮减性筛选发现，随机挑选的30个单克隆中，其中ZS-9对HepG2具有较高亲和力，氨基酸测序结果表明，该序列与美国国立生物技术信息中心（NCBI）GenBankDNA序列数据库和SwissProt蛋白数据库中的已知基因和蛋白无同源性，而且，国内外文献均未见报道，表明笔者筛选到一新的肝癌相关抗原的配体。结论利用噬菌体随机12肽库成功筛选到与肝癌细胞HepG2具有较高亲和力的多肽，为筛选鉴定新的肝癌特异的标志物奠定工作基础，也为肝癌的早期诊断和靶向治疗进一步研发确定了靶标。Objective To obtain short peptides which specifically binds to HepG2 cell line from 12 peptide libraries, and lay foundation for screening and identifying the new liver cancer markers for early diagnosis and treatment of liver cancer. Methods The liver cancer cell line HepG2 was used as the antigen and LO-2 as the absorber cells for subtraction biopanning from a phage display peptide library at 37%. The positive phage clones were identified by cell enzyrne-linked immunosorbentassay （ELISA）, and the identity of DNA sequence and amino acids were analyzed. Results After 3 rounds of screening, 30 phage clones were identified by ELISA, ZS-9 of them bind to the HepG2 specifically. The amino acid sequence was blast in NCBI and Swiss Prot, the results show that the sequence has not identity with the known genes and proteins in the database, and the sequence was not reported in literature. All these show that we had discovered several novel liver cancer associated antigen ligand. Conclusion Several candidate peptide binding to liver cancer specifically have been selected by phage display technology, providing the potential uses for early diagnosis of liver cancer or targeted therapy for liver cancer.

关 键 词：癌 肝细胞 噬菌体肽库 多肽 HEPG2 减性筛选 

分 类 号：R735.7[医药卫生—肿瘤]