关键词: 内皮抑素;基因;真核表达

    中图号:Q75

    摘 要: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法将大鼠血清白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

    FU Luo-An，ZHANG Xiang，WU Jing-Wen，GAO Da-Kuan，YANG Li-Sun，QU Yan

    Institute of Neurosurgery of Chinese PLA，Xijing Hospital，Fourth Military Medical University，Xi'an710033，China

    Keywords:endostatin;gene;eukaryotic expression

    Abstract:AIM To construct the expression vector of endo-statin gene and transfect it into C6cell，and to discuss its an-tiangiogenesis 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 pcDNA3to construct pcDNA-SE.The vector was transfected into C6glioma cells by lipofectamine and the positive clone was screened by G418.The activity of endostatin protein expressed by the C6cells was examined by immunohistochemistry and the cell proliferation assay.RESULTS The eukaryotic expression vector pcDNA-SE was successfully constructed and transfect-ed into C6cells.The cells expressed the endostatin protein which could inhibit the endotheliocyte proliferation.CONCLUSION Endostatin is a potent angiogenesis in-hibitor.The experiment lays a foundation for following ex-periments on antiangiogenesis gene therapy of tumor. ......