【摘要】 目的在人脐静脉内皮细胞(HUVECs)以及鸡胚中来研究抗碱性成纤维细胞生长因子(bFGF)单抗对血管新生的作用。方法制备3种腹水型bFGF单抗MabF7,MabF10,MabF12。CCK-8法检测bFGF单抗对HUVECs细胞增殖的影响;Transwell小室研究bFGF单抗对HUVECs迁移的影响;ECM gel检测其对HUVEC体外成管的影响;并研究其体内对鸡胚尿囊膜(CAM)血管新生作用。结果 MabF7,MabF10,MabF12均可中和bFGF的活性;3株单抗均可抑制内皮细胞迁移过程,无抗体组,对照抗体组,MabF7,MabF10,MabF12组细胞迁移率分别为100%,106.25%±7.89%,69.50%±6.86%,74.00%±4.16%,67.75%±3.30%;3株单抗均可抑制内皮细胞成管过程,无抗体组,对照抗体组,MabF7,MabF10,MabF12组管状结构形成率分别为:100%,105.93%±3.85%,56.53%±4.35%,29.23%±6.45%,12.77%±2.67%;计数尿囊膜上加药滤纸周围呈放射状的血管条数,bFGF组,bFGF+MabF7组,bFGF+MabF10组,bFGF+MabF12呈放射状的血管条数分别为15±0.82,7.5±1.29,13.5±3.10,8.5±0.58。结论 bFGF单抗对HUVECs的增殖、迁移、成管以及鸡胚尿囊膜血管新生均有抑制作用,从而为具有抗肿瘤作用的抗体药物的研发奠定基础。更多还原

【Abstract】 The aim is to research the antiangiogenic activities of anti-bFGF monoclonal antibody in human umbilical vein endothelial cell(HUVECs) and chicken embryo allantois membrane(CAM).Three strains of anti-bFGF monoclonal antibodies,MabF7,MabF10,MabF12,were prepared and purified.We examined the HUVEC proliferation,migration,and tube formation by Cell Counting Kit(CCK-8),Transwell Cab,and ECM gel,respectively.The results showed that MabF7,MabF10,MabF12 could neutralize bFGF and suppress the proliferation,migration and tube formation of the HUVEC cell in vitro.The migration ratio of medium control,antibody control,and anti-bFGF antibodies(MabF7,MabF10,MabF12) were respectively 100%,106.25%±7.89%,69.50%±6.86%,74.00%±4.16%,67.75%±3.30%;the tube formation ratio were respectively 100%,105.93%±3.85%,56.53%±4.35%,29.23%±6.45%,12.77%±2.67%.Also,the anti-bFGF antibodies can inhibit the angiogenesis in CAM.The number of the radiated micrangium in bFGF control,bFGF+MabF7 group,bFGF+MabF10 group,bFGF+MabF12 were respectively 15±0.82,7.5±1.29,13.5±3.10,8.5±0.58.The results concluded that the anti-bFGF monoclonal antibody can inhibit the proliferation,migration,and tube formation of blood endothelium cells in vitro,and the anti-bFGF antibody can also inhibit the angiognensis in CAM,which will promote the research of antitumor medicine.更多还原