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目的 观察血管内皮生长因子受体 (VEGFR)结合肽 -颗粒酶 B(Gra B)融合蛋白抗血管生成的作用 .方法 抽提人外周血淋巴细胞总 RNA,进行 RT- PCR克隆 Gra B c DNA;抽提 L o Vo细胞总 RNA,进行 RT- PCR克隆 VEGFR结合肽c DNA,用限制性酶切和 DNA测序进行鉴定 ;通过聚丙烯酰胺凝胶电泳 (SDS- PAGE)分析表达产物 .表达产物经亲和层析纯化后 ,以人血管内皮细胞 (ECV30 4 )和鸡胚尿囊膜血管测定其生物学活性 .结果 表达产物以可溶性分子形式存在于菌体中 ,具有良好的抗原性和特异性 ,且具有抑制血管内皮细胞增殖及破坏鸡胚尿囊膜血管形成的活性 .结论 VEG-FR结合肽 - Gra B融合蛋白具有抑制血管生成的功能 ,在肿瘤生物靶向治疗中有一定潜在价值
Objective To investigate the anti-angiogenic effect of VEGFR binding peptide-Granzyme B (GraB) fusion protein.Methods Total RNA was extracted from peripheral blood lymphocytes and cloned by PCR. Total RNA was extracted from Lovo cells and cloned by RT-PCR. The VEGFR-binding peptide c DNA was identified by restriction enzyme digestion and DNA sequencing. The expressed product was analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) After purification by affinity chromatography, the biological activity was determined by human vascular endothelial cells (ECV30 4) and chick chorioallantoic membrane.Results The expressed product was soluble in the form of soluble molecules and had good antigenicity and specificity , And had the activity of inhibiting the proliferation of vascular endothelial cells and destroying the angiogenesis of chick chorioallantoic membrane.CONCLUSION VEG-FR binding peptide-Gra B fusion protein has the function of inhibiting angiogenesis and has certain potential value in tumor bio-targeted therapy