全细胞差减筛选法筛选与人卵巢癌细胞特异性结合的短肽  

Screening of short peptides that bind specifically to ovarian cancer cell line through whole-cell subtraction biopanning

作　　者：周聪[1] 康佳丽[1] 王小霞[1] 聂妙玲[1] 蒋文燕[1] 

机构地区：[1]广州市第一人民医院妇产科,广东广州510180

出　　处：《细胞与分子免疫学杂志》2013年第10期1098-1101,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：广州市科学技术局(11A53150713);广东省自然科学基金(S2012010009341)

摘　　要：目的全细胞差减法筛选人源性卵巢癌HO8910细胞株特异性结合短肽,为卵巢癌的靶向药物治疗提供理想的载体。方法以人卵巢癌HO8910细胞为靶细胞,人正常卵巢上皮细胞为吸附细胞,对噬菌体随机环7肽库进行4轮差减筛选;ELISA验证阳性噬菌体克隆;对获得的阳性克隆进行DNA测序及生物信息学分析,采用免疫荧光细胞化学方法鉴定噬菌体阳性克隆(phage1)与HO8910细胞的特异性结合。结果经过4轮筛选后,噬菌体在靶细胞HO8910上出现了明显的富集现象;ELISA对20个随机挑选的克隆进行鉴定,12个可与HO8910特异性结合;DNA测序后得到一段与卵巢癌细胞特异性结合的7肽(SWQIGGN),免疫荧光细胞化学结果显示phage1能特异性结合HO8910细胞。结论采用全细胞差减法成功筛选到与卵巢癌细胞特异性结合的7肽。Objective To obtain short peptides that bind specifically to ovarian carcinoma cell line HO8910 by whole-cell subtraction biopanning as an ideal vector for targeted drug delivery in ovarian cancer therapy. Methods With the HO8910 ovarian carcinoma cells as the target cells and human normal ovarian epithelial cells as the adherent cells, 4 rounds of panning from a PH. D-CTC phage-display peptide library were carried out. Individual phage clones were selected and identified by ELISA. Positive phage clones were characterized with DNA sequencing and bioinformatics analysis. The specific binding of the positive phage clones to HO8910 cells was tested with immunofluorescence cytochemistry. Results After 4 rounds of biopanning, phage clones showed preferential binding to the target cells. ELISA identified 12 positve phage clones from 20 randomly selected ones. Heptapentide （SWQIGGN） that bound specifically to ovarian cancer cells was obtained by DNA sequencing. The results of immunofluorescence cytochemistry indicated that phage1 could be specifically bound to HO8910 cells. Conclusion By means of whole-cell subtraction biopanning, we found a novel heptapentide which was able to bind specifically to ovarian cancer cells.

关 键 词：卵巢癌 噬菌体展示 全细胞 差减筛选 靶向性 

分 类 号：R737.31[医药卫生—肿瘤] R392.33[医药卫生—临床医学]