【摘要】 目的利用噬菌体随机肽库筛选特异性靶向人肝癌细胞的短肽,为肝癌的靶向治疗提供理论依据。方法以人肝癌细胞HepG2为靶细胞,对噬菌体随机十二肽库进行3轮筛选。建立HepG2荷瘤裸鼠实验动物模型,并在裸鼠体内对经过体外3轮筛选的肽库再进行1轮筛选。随机挑取30个阳性噬菌体克隆进行序列测定及同源性分析。通过免疫细胞化学染色、免疫组织化学染色鉴定噬菌体展示肽的特异性。结果经过3轮体外筛选和1轮动物体内筛选,噬菌体在靶细胞HepG2上出现明显富集,随机挑选克隆测序结果表明十二肽VRKRSECLGAHD出现次数最多,免疫细胞化学染色、免疫组织化学染色进一步证明该噬菌体能特异结合于肝癌细胞。结论筛选得到的短肽能够特异性与肝癌细胞结合,为进一步研制用于治疗肝癌的高靶向性药物奠定实验基础。更多还原

【Abstract】 Objective To screen the peptides that bind specifically to the hepatoma cells using phage display random peptides library. Methods Three rounds of panning were conducted in vitro targeting HepG2 cell lines. In nude mice bearing HepG2 tumor, one round of panning was conducted, and 30 phage clones were randomly selected for sequence analysis to identify the consensus sequence. Immunocytochemistry and immunohistochemistry were performed to determine the specificity of the phages to the hepatoma cells. Results After three rounds of panning in vitro and one round of panning in vivo, the phages binding to HepG2 cells were enriched. Sequence analysis of the randomly selected clones identified the peptide sequence VRKRSECLGAHD as the most frequent sequence. Immunocytochemistry and immunohistochemistry confirmed the specificity of the phage binding to the hepatoma cells. Conclusions Specific peptides against hepatoma cells can be obtained from a phagedisplay peptide library, which provides an experimental basis for developing therapeutic agents targeting hepatoma cells.更多还原