肝癌特异性黏附肽的鉴定和分析  

Identification and localization of hepatocarcinoma-specific adhesion peptide

作　　者：吴淼[1] 杜冰[1] 汪磊[1] 周忠良[1] 钱旻[1] 于静[1] 

机构地区：[1]华东师范大学生命科学学院免疫学实验室,上海200062

出　　处：《现代免疫学》2006年第2期143-146,共4页Current Immunology

基　　金：上海市教委曙光计划资助项目(02SG21);上海市科技发展基金重点资助项目(04JC14033);国家自然科学基金资助项目(30440039);上海市科委纳米专项基金资助项目(0352nm113)

摘　　要：利用体内噬菌体随机肽库技术筛选出肝癌特异性噬菌体肽A54,随后用化学合成法在体外合成该多肽,并标记荧光素和生物素,在组织和细胞水平对该肽进行了体内外肝癌特异性的鉴定及其抗原定位。将生物素(Biotin)标记的A54肽通过尾缘静脉注射入荷瘤裸鼠体内,通过免疫组化方法,检测A54肽与肝癌组织特异性结合的情况;并用免疫荧光细胞化学技术,对荧光素(FAM)标记A54肽进行了细胞水平的肝肿瘤特异性鉴定及抗原定位。体内外检测结果表明,A54肽仍保留了特异性识别肝癌组织的特性,且结合于肝癌细胞的膜表面。实验证明,化学合成的多肽,在体内仍然具有靶向肝癌细胞膜表面的作用,这进一步证明了筛选出来的目标噬菌体克隆其特异性靶向作用确实是由其表面插入的小分子多肽所介导的,为肝癌特异性导向药物载体的研究提供了科学依据。The hepatocarcinoma specific adhesin peptide A54 was screened by means of the phage display technology in vivo and this peptide was chemically synthesized in vitro. To confirm its potential elincal application in the targeted therapy of liver cancers, the i dentification and localization of this peptide were performed by introducing the biotin-labeled A54 peptide intravenously through tail vein into the tumor-bearing nude mice and examining the specific binding of A54 peptide with hepatocarcinoma tissues by immuno-histochemical methods. The liver tumor-specific identification and antigen localization of the fluorescin-labeled A54 peptide were performed under cellular level. The experimental results showed that the property of A54 peptide to recognize specifically the liver cancer tissues was demonstrated and its binding with tumor tissues occurred on the surface membrane of liver cancer cells. It was also demonstrated that the chemically synthesized peptide still had the property to target on the surface membrane of liver cancer cells in vivo, thus further indicating that the specific targeting effect of the objective phage clone was mediated by the small molecule of peptides insertd into the cell membrane. It is evident that the A54 peptide would be an excel lent candidate for the targeting therapy to hepatocarcinoma.

关 键 词：噬菌体肽 生物素标记肽 荧光素标记肽 免疫组化 体内靶向作用 抗原定位 

分 类 号：Q782[生物学—分子生物学]