重组人内皮抑素的表达纯化及对内皮细胞增殖的影响

EXPRESSION AND PURIFICATION OF RECOMBINANT HUMAN ENDOSTA TIN AND ITS EFFECT ON ENDOTHELIAL CELL PROLIFERATION

目的探讨融合表达的重组人内皮抑素对人脐带静脉血管内皮细胞(ECV 304)增殖的影响。方法将pTRX-endo质粒转化大肠杆菌BL21(DE3),IPTG诱导表达,经纯化后加入ECV 304细胞培养,MTT法检测对内皮细胞增殖的影响。结果经SDS-PAGE分析,出现特异性蛋白质条带,纯化后得到纯度在95%以上的重组人内皮抑素蛋白,明显抑制ECV 304的增殖。结论内皮抑素可明显抑制内皮细胞增殖,存在剂量效应关系,本实验为探索内皮抑素抑制血管生成的机制及应用研究打下基础。

Objective To investigate the effect of recombinant human endostatin expressed in E. coli on the proliferation of ECV304 cells. Methods pTRX- endo plasmid was expressed in E. coli BL21 （DE3）. Endostatin was expressed in E. coli by IPTG inducement, ECV304 cell were added to it for culture after purification, and its prolification effect was analyzed by MTT. Results SDS - PAGE analysis showed a specific protein strip, the purity of recombinant human endostatin was over 95 % after purification. Endostatin inhibited obviously the proliferation of ECV304 cells. Conclusion Recombinant human endostatin may antagonize the proliferation of endothelial cell. The experiment lays foundation for the research of antiangiogenesis and application of endostatin.