抗人肝癌血管内皮功能性单抗的筛选与鉴定  

Screen and identify of anti-human liver cancer vascular endothelium functional monoclonal antibody

作　　者：孙力超[1] 朱小飞[2] 遇珑[1] 孙立新[1] 杨治华[1] 冉宇靓[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤研究所分子肿瘤学国家重点实验室,北京100021 [2]新乡医学院医学检验系免疫研究中心,河南新乡453003

出　　处：《中华肿瘤防治杂志》2011年第11期829-832,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家重点基础研究发展计划(973)资助项目(2009CB521804)

摘　　要：目的：筛选、鉴定抗人肝癌血管内皮功能性单抗，为治疗肝癌提供靶向治疗剂，并为分离获得肝癌相关的分子靶标打下基础。方法：采用活细胞荧光、MTT细胞增殖实验、成管实验和动物体内治疗实验，筛选鉴定抑制肝癌内皮细胞的功能性单抗。结果：从能与肝癌内皮细胞膜反应的119株克隆中，筛选出16株单抗显著地抑制肝癌内皮增殖其抑制率达21％～46％，46株能显著抑制肝癌内皮细胞的成管，3株单抗1F9、1285和1811能显著抑制肝癌移植瘤的生长抑制率分别为50．8％、48．7％和47．0％。选择1株体内抑瘤效果最好的1811单抗，通过抗体抗原亲和层析法进行纯化，并对纯化的单抗的相关抗原蛋白进行免疫组织化学实验鉴定，结果显示1811抗原在肝癌血管组织较高的表达而在正常肝血管组织极少表达。蛋白质印迹法显示其抗原相对分子质量约46×10^3。结论：采用大容量功能性抗体库技术成功获得了多株具有抑制肝癌内皮恶性生物学行为的功能性单抗，体内外抑制肺癌的生长，具有成为肺癌靶向治疗剂的潜力。其中1株可能是一个靶向治疗肝癌的新靶位。OBJECTIVE： To screen and identify functional monoclonal antibodies against human liver cancer vascular endothelium （HLCVE）, so as to provide a candidate antibody drug and molecule target for the anti-liver cancer therapy. METHODS： The inhibitive monoclonal antibody were studied by immunofluores cence, ELISA, cell proliferation assay,tube formation assay and in vivo tumor treatment experiments. The corresponding antigen was identified by Western blotting. RESULTS： From the 119 monoclonal antibodies, which could recognize cell surface protein of HL- CVE, 16 monoclonal antibodies were selected which could inhibited HLCVE proliferation in vivo; The inhibition rate was 21G 46G. Forty-six monoclonal antibodies could supressed HLCVE tube formation. 1F9,12B5 and 1Bll could inhibit the growth of liver tumor and the inhibition rates were 50.8G,48.7% and 47.0G. By IHC, the results indicate that 1Bll mAb, which could significiently inhibit tumor frowth, associated antigen protein express only in liver tumor blood vessel but seldom found in normal liver blood vessel. Western blotting revealed that the molecular weight of the antigen of 1Bll antibody was about 46 X 103. CONCLU- SIONS： Several clones of inhibitory monoclonal antibodies were successfully obtained which can supress the function of HLCVE in vitro and in vivo. 1Bll monoclonal antibody can inhibit liver tumor growth in vitro and in vivo ; which may become a candidate drug for targeted liver cancer treatment.

关 键 词：肝肿瘤 内皮细胞 抗体 单克隆 

分 类 号：R735.7[医药卫生—肿瘤]