Endostatin基因真核表达载体的构建及活性鉴定

目的：构建endostatin基因的真核表达载体，并将其转染C6脑胶质瘤 细胞，探讨其体外抑制血管内皮细胞生长的活性。方法：利用PCR将大鼠血清蛋白分泌信号连接在endostatin基因的碳末端，构建成真核表达载体pcDNA3-Sendostatin，利用Lipofectamine法将其转染C6脑胶质瘤细胞，采用细胞增殖实验进行endostatin表达蛋白体外活性鉴定。结果：成功构建了endostatin基因真核表达载体，转染该载体的C6脑胶质瘤细胞可分泌表达抑制血管内皮细胞增生的endostatin蛋白。结论：endostatin是一种有效的血管生成抑制剂，可进一步用于在体肿瘤的抗血管生成治疗。

Construction of the eukaryotic expression vector of endostatin gene and identification of its activity

AIM To construct the expression vector of endostatin gene and transfect it into C6 cell, and to discuss its antiangiogenesis activity. METHODS The rat serum albumin secretive signal was linked to the endostatin gene C terminal by PCR, this fused gene was then inserted into polylinker sites of eukaryotic expression vector pcDNA3 to construct pcDNA SE. The vector was transfected into C6 glioma cells by lipofectamine and the positive clone was screened by G418. The activity of endostatin protein expressed by the C6 cells was examined by immunohistochemistry and the cellproliferation assay. RESULTS The eukaryotic expression vector pcDNA SE was successfully constructed and transfected into C6 cells. The cells expressed the endostatin protein which could inhibit the endotheliocyte proliferation. CONCLUSION Endostatin is a potent angiogenesis inhibitor. The experiment lays a foundation for following experiments on antiangiogenesis gene therapy of tumor.