C6细胞表达的人Angiostatin Kringle(1-3)体外抑制血管内皮细胞生长的研究

为研究人AngiostatinKringle( 1 3) [AK( 1 3) ]抑制血管内皮细胞生长的活性 ,利用脂质体法将带有大鼠血清白蛋白分泌信号的人AK( 1 3)基因导入大鼠C6胶质瘤细胞 ,并用G41 8筛选获得目的细胞株。采用电镜、流式细胞术、免疫组化和Westernblot等方法 ,检测转染前后细胞超微结构、细胞周期及AK( 1 3)蛋白表达情况 ,并用人脐静脉内皮细胞增殖实验检测目的细胞株表达的AK( 1 3)蛋白抑制血管内皮细胞生长的活性。结果表明 ,成功转染人AK( 1 3)基因的大鼠C6胶质瘤细胞可稳定表达AK( 1 3)蛋白 ,且具有抗血管内皮细胞增殖活性的良好作用 ,为进一步进行体内抗血管生成基因治疗奠定了基础。

XPRESION OF HUMAN ANGIOSTATIN KRINGLE(1-3) IN C6 CELL SUPPRESSESENDOTHELIAL CELL PROLIFERATION IN VITRO

To research anti angiogenesis activity of, human AK(1 3)[angiostatin kringle(1 3)],rat serum albumin signal pipeptide was transfected into C6 cell utilizing lipofectamine. Electron microscope, flow cytometry, immunohistochemistry and Western blot were used to detect the ultramicrostructure, cell cycle and expression of AK(1 3) of C6 cell transfected and untransfected. The anti angiogenesis activity of AK(1 3) expressed by C6 cell was identified with human umbilical vein endothelial cell proliferation assay. The results indicate that C6 cell succsessfully transfected with AK(1 3) gene can stably express active AK(1 3) which potently inhibits endothelial cell proliferation. The study provides a promise for the anti angiosgenesis genetherapy in vivo.