摘要
目的 研究血管内皮生长因子 (VEGF)受体flt 1胞外区基因抗肿瘤新生血管生成的作用。 方法 采用脂质体转染技术将含有flt 1胞外区前三个IgG样结构区域的真核表达质粒 pcD NA3.1/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PCR和RT PCR鉴定。对裸鼠人膀胱癌组织切片进行免疫组化染色 ,定量分析微血管密度。 结果 PCR和RT PCR结果显示重组质粒转入EJ细胞及在转录水平表达 ;免疫组化的微血管密度实验组明显低于阴性对照组。 结论 阻断VEGF/flt 1信号传导途径能够抑制肿瘤新生血管的形成 。
Objective To verify whether the extracellular domain of flt 1 has anti angionesis activity in vivo. Methods A recombinant plasmid pcDNA 3.1/flt 1 n3 was transfected into the human bladder carcinoma EJ cell by lipofectamin, which was constructed by inserting the N terminal first three IgG like domain of VEGF receptor flt 1 (flt 1 n3 ) into eukaryotic expression vector pcDNA 3.1. Results The rflt 1 was functionally expressed in stably transfected EJ and the product secreted in the medium could be specifically bind to rhVEGF165.The result showed that the bladder cancer transfected with rflt 1 had a lower microvessle density than the control. Conclusions It is proved that the expressed rflt 1 n3 can inhibit tumor growth and angiogenesis in nude mice model.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2001年第8期460-463,共4页
Chinese Journal of Urology
基金
国家 8 63高技术规划基金资助 (10 2 0 8 0 1 0 3 )
