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Preliminary screening and identification of the hepatocarcinoma cell-binding peptide
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NUCLEAR MEDICINE
- Issue:
- 6
- DOI:
- 10.3760/cma.j.issn.2095-2848.2004.06.006
- Key Word:
- 肝肿瘤;肿瘤细胞,培养的;肽类;基因疗法
Abstract： 目的探讨利用噬菌体随机肽库筛选肝癌细胞特异性结合多肽或肝癌细胞优势结合多肽的可行性.方法①利用噬菌体随机十二肽库,对人肝癌细胞系HepG2和人正常肝细胞系L02进行4轮差异筛选、富集,获得与肝癌细胞亲和的噬菌体克隆.②用酶联免疫吸附测定(ELISA)法测定其与肝癌细胞、正常肝细胞的亲和力,进一步筛选出高度亲和肝癌细胞的特异性阳性噬菌体克隆.比较其与其他肿瘤细胞的亲和力.③提取阳性噬菌体DNA并进行DNA序列测定,获得与肝癌细胞亲和的多肽序列.④用免疫荧光共聚焦成像进一步确定展示阳性多肽序列的噬菌体与肝癌细胞的结合部位.⑤根据筛选的多肽序列合成十六肽WH16,利用竞争抑制实验鉴定其与肝癌细胞的结合.结果筛选及ELISA结果示得到与肝癌细胞特异性结合并内化入肝癌细胞的噬菌体克隆,测序结果示56.67%的被检噬菌体展示相同的多肽序列FLJEPHLMDTSM,推测FLEP是肝癌细胞特异性结合肽的基序.免疫荧光共聚焦成像进一步确定展示序列FLLEPHLMDTSM的噬菌体与肝癌细胞特异性结合并内化入细胞内.竞争抑制实验示WH16能有效抑制展示序列FLLEPHLMDTSM的噬菌体克隆与肝癌细胞的结合.结论利用噬菌体随机肽库获得能与肝癌细胞特异性结合的多肽序列FLLE-PHLMDTSM,据此合成的十六肽WH16与肝癌细胞有亲和力.
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