中文摘要：

设计并构建一种含VEGF和GRP抗原表位的表达载体pET28a—VEGFI-M2-GRP质粒，将其转化至大肠杆菌中，并对重组菌进行乳糖诱导表达，超声破碎，包涵体经洗涤、溶解、透析复性、离子交换层析等方法进行分离纯化后得到目的蛋白VEGFⅡ／GRP，Westernblot鉴定。建立前列腺癌RM-1细胞的C57BL／6J小鼠皮下移植瘤模型，以VEGFⅡ／GRP作为疫苗进行免疫。观察荷瘤小鼠的肿瘤生长情况计算抑瘤率，比较各组血管生成数以及EHSA检测抗VEGF抗体浓度，并研究该蛋白疫苗的抗肿瘤生长作用和抗血管生成作用。结果显示：EHSA结果表明小鼠血清中抗VEGF抗体比NS组高（P〈0．05），重组蛋白VG组与NS组相比抗血管作用显著（P〈0．05）。初步显示构建的重组蛋白有抑制肿瘤血管生长的作用。

英文摘要：

An expression vector pET-28a-VEGF I -M2-GRP plasmid containing the VEGF and GRP antigen epitopes was designed and constructed. After being expressed in recombined strains induced by lactose, the fusion protein was lysised by ultrasonic. The resulting inclusion body was washed, dissolved, and dialysised for renaturation and purified by ion-exchange chromatograph to get the target protein. VEGF 11/GRP was identified by Western blot. The C57BL/6J mice subcutaneous tumor model of prostate cancer cell line RM-lwas established to receive treatments with VEGF II/GRP vaccine. The tumor growth, the blood vessel densities and the concentration of anti-VEGF antibody were examined to assess the effect of anti-angiogenesis and anti-tumor. ELISA showed that the concentration of anti-VEGF antibody in the mice serum of VG group was significantly higher than that of NS group （P 〈 0.05 ）, the anti-angiogenesis effect of VG group was superior to that of NS group （P 〈 0.05 ）. Thus, the constructed fusion protein can inhibit angiogenesis.