MDA-MB-231乳腺癌细胞特异性多肽的初步筛选

Screen peptides specifically bonding to MDA-MB-231 breast cancer cells

目的 利用噬菌体展示技术,从噬菌体随机十二肽库中筛选出能够特异性结合MDA-MB-231乳腺癌细胞的噬菌体克隆.方法 以人正常乳腺细胞为减性筛选细胞、MDA-MB-231乳腺癌细胞为靶细胞,对噬菌体随机十二肽库进行筛选,挑取富集后的阳性单克隆噬菌体,酶联免疫吸附试验（ELISA）及DAB染色鉴定阳性噬菌体的特异性及亲和力.结果 经过3轮筛选,噬菌体得到约113倍的富集,随机挑选11株单克隆噬菌体,ELISA显示8号噬菌体单克隆对乳腺癌细胞的亲和力是对照的6.5倍,DAB鉴定亦显示其对乳腺癌细胞的特异性及亲和力最高,命名为LK-8.结论 利用噬菌体筛选技术成功筛选出能够特异性结合MDA-MB-231乳腺癌细胞的特异性噬菌体单克隆LK-8,可为进一步合成特异性多肽用于早期诊断和靶向治疗乳腺癌奠定基础.

Objective To screen phage clones which can specific binding to MDA-MB-231 breast cancer cells from phage random 12 peptide library by phage display technology.Methods Agminated positive monoclonal phage was selected from phage random 12 peptide library,using normal mammary gland cells as subtract screening cells and MDA-MB-231 breast cancer cells as target cells.The specificity and affinity of the positive clones were identified by enzyme linked immunosorbent assay （ELISA） and DAB dyeing.Results Eleven clones were chosen after 3 rounds of screening.ELISA showed the affinity to breast cancer cells of NO.8 phage （named LK-8） was 6.5 times higher than that in control group,immunohistochemistry also showed NO.8 phage had a high specificity bonding to breast cancer cells.Conclusion A specific phage （LK-8）bonding to MDA-MB-231 breast cancer cells is screened successfully using phage display technology,and it lays the foundation for compounding specific polypeptide to diagnose and treat breast cancer.