重组人分泌型endostatin的纯化及其抑癌活性探讨

目的：纯化毕赤酵母菌GSll5株表达的分泌型人endostatin蛋白，并对其抑癌活性进行研究。方法：利用SepharoseG—25及Heparin亲和层析柱纯化重组蛋白，并以MTT法测定重组endostatin对人脐静脉血管内皮细胞(ECV-304)增殖的抑制活性；另以HepG2.2．15细胞注射裸鼠成瘤后，体内验证重组蛋白的抑瘤活性。结果：MTT结果显示纯化后的endostatin在体外可特异性抑制人脐静脉内皮细胞的增殖，最高抑制率为86．0％；动物实验证明其可明显抑制荷瘤动物肿瘤的生长。结论：经纯化后的重组endostatin具有较强抑癌活性，将为临床抗血管生成治疗实体瘤研究奠定基础。

Purification of Recombinant Human Secreted Endostatin and Study of Its Inhibition Effect on Tumor

Objective:Purify recombinant human secreted endostatin expressed in Pichia Pastoris(Pp)GS115strain,and study its inhibition effect on tumor.Methods:Sepharose G25and heparin affini ty chromatography column were used to purify recombinant human secreted en-do statin.MTT assay was used to detect the inhibitory effect on proliferation of e ndothelial cell line.Furthermore,nude mice bearing HepG2.2.15cells were injected with recombinant protein,the size of tumor was measured2weeks later .Results:Purified endostatin may inhibit the prolif-eration of endothelial ce ll in vitro,the highest inhibitory rate was86%.It can inhibit tumor to grow and induce tumor cell to apoptosis in vivo of animal.Conclusion:Purified endo statin has strong-ly inhibitory activity to tumor.This lays the foundation for the antiangiogenesis therapy of the sol-id tumor.