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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 ,延缓肿瘤的生长速度
Objective To study the antitumor neovascularization of extracellular domain of vascular endothelial growth factor (VEGF) receptor flt 1. Methods The eukaryotic expression plasmid pcD NA3.1 / KDRn7 containing the first three IgG-like structural regions of extracellular domain of flt 1 was transfected into human bladder cancer EJ cells by lipofection. The recombinant protein was stably expressed by G418 The cell clones were screened by solid-phase binding assay to obtain the cell clones expressing the specific protein binding with VEGF. The positive cell clones were identified by PCR and RT PCR. Immunohistochemical staining of human bladder cancer tissue sections of nude mice to quantitatively analyze the microvessel density. Results PCR and RT PCR results showed that the recombinant plasmids were transfected into EJ cells and expressed at the transcriptional level. The experimental group with immunohistochemical microvessel density was significantly lower than the negative control group. Conclusion Blocking the VEGF / flt1 signaling pathway can inhibit tumor angiogenesis and delay the growth of tumor