Establishment and Characterization of a Targeting Tumor Vesseles System for Inducing Thrombosis

目的建立一种新型的靶向高效诱发肿瘤血管血栓栓塞体系磁流体-神经纤维网蛋白1抗体A6-链霉亲和素:生物素-短截型组织因子(Mf-A6-SA:b-TTf)。方法化学交联技术制备神经纤维网蛋白1抗体A6-链霉亲和素、磁流体-神经纤维网蛋白1抗体A6-链霉亲和素和生物素-短截型组织因子交联物,凝血因子X活化实验鉴定复合体系活化凝血因子X的活力,荧光显微镜技术和普鲁士蓝染色法同时观察施加外界磁场后复合体系的靶向作用,凝血实验直接观察复合体系引入链霉亲和素:生物素的生物放大效应,体内生物分布实验观察复合体系的安全性。结果成功制备磁流体-神经纤维网蛋白1抗体A6-链霉亲和素及生物素-短截型组织因子,磁流体-神经纤维网蛋白1抗体A6-链霉亲和素:生物素-短截型组织因子体系保留有高效激活凝血因子X的活性,与靶点的结合具有靶向性及高效富集性,体内实验证实能安全有效诱发肿瘤血管栓塞。结论成功制备的具有靶向诱发肿瘤血管血栓栓塞体系磁流体-神经纤维网蛋白1抗体A6-链霉亲和素:生物素-短截型组织因子为进一步探索肿瘤血管的靶向治疗奠定基础。OBJECTIVE To establish a novel targeting tumor vessels system MF-A6-SA:B-tTF for efficiently inducing thrombosis.METHODS Chemical cross-linking technique was used to prepare a cross-linking agent of the A6-Streptavidin( A6-SA),MFA6-SA and Biotein-tTF(B-tTF).FX coagulation assay was used to test MF-A6-SA:B-tTF system's FX activity.Fluorescence microscopy and prussian blue staining were used to simultaneously observe the targeting activity of MF-A6-SA:B-tTF with an external magnetic field.Hemagglutination was directly used to study the system' s biological amplification by SA /B.Biodistribution experiment was used to observe the toxicity of MF-A6-SA:B-tTF.RESULTS MF-A6-SA and B-tTF were successfully prepared.MF-A6-SA:B-tTF system could activate FX,inducing the blood coagulating cascade powerfully.MF-A6-SA:B-tTF could be accumulated to the desired target area with targeting and induce thrombosis in tumor blood vessels in vitro and in vivo.CONCLUSION The double targeting tumor vessels system MF-A6-SA:B-tTF maybe provide a basis for developing the tumor blood vessels targeting therapy.国家自然科学基金资助项目(30973485;81172970); 国家级大学生创新性实验项目(201210384134