重组色素上皮细胞衍生因子的制备及其对脑胶质瘤血管新生的影响  

The acquisition of recombinant pigment epithelium-derived factor and the effect on the angiogenesis of gliomas

作　　者：张弢[1] 关明[1] 吕元[1] 

机构地区：[1]复旦大学上海医学院附属华山医院检验医学中心,上海200040

出　　处：《中华检验医学杂志》2005年第5期483-485,共3页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目 ( 30300360 );高教部博士点基金资助项目(20030246060);上海市医学重点学科项目;上海市青年科技启明星计划资助项目(04QMX1408)

摘　　要：目的　用大肠杆菌表达色素上皮细胞衍生因子(PEDF),及评价其在脑胶质瘤血管新生中的作用。方法　构建pET41 /PEDF表达载体,用金属亲和层析(HisTag)和阴离子交换层析纯化重组蛋白,并以管腔形成试验验证重组蛋白活性。同时,采用酶联免疫吸附试验(ELISA)测定血管内皮细胞生长因子(VEGF)与血小板反应素1 (TSP- 1)的变化。结果　用基因工程手段获得终产量为3 8mg/L,可抑制内皮细胞管腔形成的PEDF蛋白; PEDF可下调VEGF(减少1 8倍)和上调TSP -1(增高5 3倍)的表达。结论　PEDF可能是抑制脑胶质瘤新生血管形成的主要因素。Objective To express the pigment epithelium-derived factor (PEDF) in E.coli. and to assess its role in angiogenesis of gliomas.Methods The expression plasmid pET41/PEDF was constructed and the protein was purified by metal affinity tag (His Tag ) and anion-exchange chromatography. Tube formation was used for evaluating its biological activity. The changes of vascular enthothelial growth factor (VEGF) and thrombospondin-1 (TSP-1) were evaluated by the enzyme-linked immunosorbent assay.Results The high purified and inhibitory-actived PEDF proteins were acquired and PEDF had the function of decreasing the VEGF 1.8 times as well as increasing the TSP-1 5.3 times.Conclusions It is possible that PEDF expression is a potent factor for the inhibition angiogenesis in gliomas.

关 键 词：色素上皮细胞衍生因子 脑胶质瘤 血管新生 血管内皮细胞生长因子(VEGF) 酶联免疫吸附试验(ELISA) 制备 大肠杆菌表达 PEDF 血小板反应素 新生血管形成 管腔形成 阴离子交换 表达载体 亲和层析 重组蛋白 层析纯化 蛋白活性 

分 类 号：R739.4[医药卫生—肿瘤]