Extracellular domain of kinase domain region mediated by adeno-associated virus inhibits growth and angiogenesis of bladder cancer in Balb-c mice

腺伴随病毒介导人血管内皮生长因子受体KDR胞外区基因对裸鼠人膀胱癌生长及血管生成的影响(英文)

OBJECTIVE: To verify whether the extracellular domain of kinase domain region (KDR) has anti-angiogenesis activity in vivo. METHODS: cDNA was cloned into adeno-associated virus (AAV) vector pSNAV and transfected to baby hamster kidney (BHK) cells. Recombinant AAV was obtained from the cell culture supernatant after adding helper virus. Recombinant AAV-infected human bladder cancer EJ cell line (EJ cells) were injected subcutaneously into Balb-c nude mice. Tumor specimens were removed from the mice, paraffin-embedded and sliced, then stained by immunohistochemistry. Microvessel density (MVD) was determined under a microscope. RESULTS: The tumor volume developed by EJ cells transfected with the extracellular domain of KDR was significantly smaller (1.70 +/- 0.18 cm(3)) compared with that in the control (5.62 +/- 0.67 cm(3)) (P

目的 从原代培养的人脐静脉血管内皮细胞 (HUVEC)提取细胞总RNA ,采用逆转录PCR(RT PCR)方法得到VEGF受体KDR全长胞外区cDNA片段 ,检验其体内抗肿瘤血管生成的作用。方法 将获得的受体基因克隆到AAV基因治疗载体pSNAV中 ,得到重组质粒pSNAV/KDR。重组质粒转染BHK细胞 ,加入辅助病毒后 ,得到表达目的蛋白的重组AAV。重组AAV表达的KDR具有与VEGF结合的活性 ,重组AAV感染人膀胱癌EJ细胞 ,皮下注射Balb c裸鼠。Ⅷ因子免疫组化染色进行微血管密度测定。结果 重组AAV感染人膀胱癌EJ细胞形成的肿瘤血管化程度明显低于对照组。