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目的 构建小鼠内皮抑素 (endostatin) c DNA的真核表达载体并将其在体外培养的脑胶质瘤细胞内表达 .方法 将带有分泌信号的小鼠 endostatin c DNA克隆入真核表达载体 pc DNA3,构建 CMV启动子控制的载体 pc DNA- s Endo,并将上述载体转染体外培养的脑胶质瘤细胞 (SHG44 ) ,采用Western Blot鉴定其表达 ,应用牛微血管内皮细胞抑制实验检测其活性 .结果 成功地构建了真核表达载体 pc DNA-s Endo并转染入脑胶质瘤细胞 ,获得的分泌型 endostatin可以明显抑制牛毛细血管内皮细胞增殖 (ED5 0 =2 0 0μg· L- 1 ) .结论 获得有表达功能活性的 endostatin真核表达载体 ,为脑胶质瘤的抗血管生成基因治疗提供了必要的条件
Objective To construct eukaryotic expression vector of mouse endostatin c DNA and express it in glioma cells cultured in vitro.Methods Mouse endostatin c DNA with secretion signal was cloned into eukaryotic expression vector pcDNA3 was constructed. The vector pcDNA-s Endo was constructed and transfected into SHG44 cells. The expression of SHG44 was detected by Western Blot. The expression of SHG44 was detected by bovine microvascular endothelial cell inhibition Activity.Results The eukaryotic expression vector pcDNA-s Endo was successfully constructed and transfected into glioma cells, and the secreted endostatin could significantly inhibit the proliferation of bovine capillary endothelial cells (ED50 = 200μg · L- 1) .Conclusion Obtaining endostatin eukaryotic expression vector with functional expression provides the necessary conditions for anti-angiogenic gene therapy of glioma