血管内皮生长因子-2 抑制剂细胞筛选模型的构建及应用

 目的 建立针对以血管内皮生长因子受体-2（vascular endothelial growth factor receptor-2， VEGFR-2）为靶标的特异及灵敏的细胞模型体系，用于酪氨酸激酶受体（receptor tyrosine kinase， RTK）抑制剂的筛选和研究。方法 构建人源VEGFR-2真核表达载体pcDNA3.1-VEGFR-2，将其转染至NIH 3T3细胞，获得稳定转染的细胞克隆，并对其进行功能学鉴定及应用；应用瞬时转染VEGFR-2的HeLa细胞，建立VEGF依赖性的受体磷酸化细胞模型及应用。结果 在无血清培养条件下，稳定转染 VEGFR-2的NIH 3T3细胞获得VEGF依赖性的细胞增殖特征；HeLa细胞经瞬时转染VEGFR-2后，可以检测出明显的VEGF依赖性的受体酪氨酸磷酸化。通过应用已知VEGFR-2抑制剂及自行合成化合物，确证以上细胞模型具有较高机制针对性的特征。结论 所构建并确证的细胞模型具有较高特异性和灵敏性，可适用于VEGFR-2和其他RTK抑制化合物的体外筛选与研究。

Construction and Application of Cell-Based Models for Screening Vascular Endothelial Growth Factor Receptor-2 Inhibitors

OBJECTIVE To construct and validate the in vitro cell-based models for the purpose of screening and investigating small molecules targeting VEGFR-2.METHODS Plasmid containing human VEGFR-2 gene was constructed and stably transfected into NIH 3T3 cells,then the function of the transfected clone cells was analyzed.HeLa cells were transiently transfected with exogenous VEGFR-2,and the VEGF-dependent tyrosine phosphorylation was examined.RESULTS The cloned cells with stable transfection of VEGFR-2 retained VEGF...