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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。
To study the activity of human Angiostatin Kringle (113) [AK (13)] in inhibiting the growth of vascular endothelial cells, the human AK (13) gene with rat serum albumin secretion signal was introduced into rat C6 by liposome method. Glioma cells were screened with G41 8 to obtain the desired cell line. Electron microscopy, flow cytometry, immunohistochemistry and Western blot were used to detect the ultrastructure, cell cycle and AK(13) protein expression before and after transfection. The expression of target cell lines was detected by human umbilical vein endothelial cell proliferation assay. The AK(13) protein inhibits the activity of vascular endothelial cell growth. The results showed that C6 glioma cells successfully transfected with human AK(13) gene can stably express AK(13) protein and have a good effect on the anti-vascular endothelial cell proliferation activity for further anti-angiogenesis in vivo. Gene therapy laid the foundation.