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的影响

目的 研究血管内皮生长因子 (VEGF)受体KDR胞外区基因对肿瘤新生血管的抑制作用。方法 采用脂质体转染技术 ,将KDR胞外区基因的真核表达质粒pcDNA 3.1/KDRn7转入人膀胱癌EJ细胞 ,用G418筛选得到稳定表达目的蛋白的细胞克隆 ,经固相结合实验筛选得到表达与VEGF特异结合的目的蛋白的细胞克隆 ,阳性细胞克隆进行了RT PCR鉴定。结果 鸡胚测活表明 ,EJ细胞表达rKDRn7的能抑制鸡胚绒毛尿囊膜的血管形成 ;表达rKDRn7的EJ细胞株对裸鼠人膀胱癌生长及血管生成明显抑制 ,实验组的微血管密度 (MVD)为 12± 4,阴性对照组 6 2± 11。结论 阻断VEGF/KDR信号传导途径能够抑制肿瘤新生血管的形成 ,延缓肿瘤的生长速度。

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

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