耐紫杉醇宫颈癌细胞株Hela特异性结合多肽的筛选及其功能鉴定  

Screening and functional identification of peptides specifically binding to Paclitaxel-resistant cervical cancer cells Hela

作　　者：李倩 左璇 邓成程 尹东锋[1] 

机构地区：[1]新疆军区总医院,乌鲁木齐830000 [2]中国人民解放军第11医院

出　　处：《山东医药》2018年第3期31-33,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81372782)

摘　　要：目的筛选与耐紫杉醇(PTX)的宫颈癌细胞系Hela(Hela/PTX)有特异性结合能力的多肽。方法以耐药宫颈癌Hela/PTX细胞为靶细胞,通过竞争结合实验,对噬菌体随机七肽库进行3轮筛选。采用ELISA法测定筛选产物中多肽与Hela/PTX细胞的亲和力(以P/N值表示)。选择P/N值较大的10条多肽进行测序,命名其中P/N值最大的一条多肽并根据其序列在体外合成此多肽。质谱检测所得多肽的分子量。细胞亲和力实验测定所得多肽与Hela/PTX、骨肉瘤细胞的亲和力。结果筛选出的多肽命名为EP-7。质谱分析显示EP-7分子量815.30D。细胞亲和力实验结果显示Hela/PTX细胞质内有绿色荧光,而骨肉瘤细胞内未观察到绿色荧光。结论成功筛选出可以与Hela/PTX细胞特异性结合的多肽EP-7。Objective To screen out the peptides specifically binding to Paclitaxel-resistant cervical cancer cells Hela（ Hela/PTX）. Methods Using drug-resistant cervical cancer Hela/PTX cells as the target cells,and the phage 7 peptide library was screened for 3 rounds by competitive binding tests. Using ELISA to determine the affinity of peptides to Hela/PTX cells（ P/N value）. Ten peptides with the high P/N value were selected to be sequenced,and we named a polypeptide with the highest P/N value and synthesized this peptide in vitro according to its sequence. The molecular weight of the peptide was determined by mass spectrometry. The affinity of the peptide to Hela/PTX and osteosarcoma cells was observed by cell affinity experiment. Results The selected peptide was named EP-7. Mass spectrometry analysis showed that the molecular weight of EP-7 was 815. 30 D. The results of cell affinity test showed that there was green fluorescence in the cytoplasm of Hela/PTX,but no green fluorescence was observed in osteosarcoma cells. Conclusion The peptide EP-7 which can bind specifically to Hela/PTX cells is successfully screened out.

关 键 词：宫颈癌 耐药宫颈癌细胞 噬菌体肽库 多肽 

分 类 号：R737.33[医药卫生—肿瘤]