卵巢癌细胞SKOV3特异性结合短肽的筛选  

Screening of specific peptides targeting to ovarian cancer cell line SKOV3

作　　者：王乐丹[1] 李文桔 胡越[1] 

机构地区：[1]温州医科大学附属第二医院妇科,浙江温州325027

出　　处：《中国妇幼保健》2015年第16期2648-2651,共4页Maternal and Child Health Care of China

基　　金：浙江省温州市科技局资助项目〔Y20140319〕

摘　　要：目的:利用噬菌体7肽库进行体外快速差减筛选(biopanning and rapid analysis of selective interactive ligands,BRASIL),从而获得卵巢癌细胞株SKOV3细胞表面特异性结合短肽。方法:以中国仓鼠卵巢细胞株CHO为吸附细胞,卵巢癌细胞株SKOV3为靶细胞,利用噬菌体展示技术联合差减筛选策略,经过连续5轮生物淘洗,随机挑选100个噬菌体单克隆进行ELISA检测,选取读数>0.6的单克隆进行DNA测序并序列分析,选择并合成相应短肽,经细胞免疫荧光验证短肽和卵巢癌细胞SKOV3结合的亲和性及特异性。结果:筛选出的短肽AFSGSLP与卵巢癌细胞株SKOV3有较高的亲和性及特异性。结论:阳性噬菌体表面展示的多肽AFSGSLP可能为卵巢癌的早期诊断和转移复发监测提供新思路。Objective： To conduct biopanning and rapid analysis of selective interactive ligands（ BRASIL） using phage peptide library,obtain specific peptide binding on the surface of ovarian cancer cell line SKOV3. Methods： BRASIL method was used to screen the7- mer phage display peptide library on Chinese hamster ovarian cell line CHO and human ovarian tumor cell line SKOV3. After five rounds of in vitro reiterative panning,100 plaque monoclones were randomly selected for ELISA,then the monoclines with the value more than 0. 6were selected for DNA sequencing. The corresponding specific peptide was synthesized,immunofluorescence assay was applied to confirm the affinity and specificity of the positive phage clone binding to SKOV3 cells. Results： The screened phage clone AFSGSLP showed high affinity and specificity combining with SKOV3 cells. Conclusion： The peptide AFSGSLP displayed on the surface of positive phage perhaps provides a new direction for early diagnosis,metastasis monitoring,and treatment of ovarian cancer.

关 键 词：卵巢癌 噬菌体展示技术 噬菌体多肽库 体外快速差减筛选技术 多肽 

分 类 号：R737.31[医药卫生—肿瘤]