重组人色素上皮衍生因子显著抑制血管内皮细胞增殖

Recombinant Human Pigment Epithelium-derived Factor Inhibits the Proliferation of the Endothelial Cells from Blood Vessels

目的通过原核表达,获得具有生物活性的人重组色素上皮衍生因子(PEDF)。方法将PEDF基因构建到GST原核表达系统中,经过表达纯化,获得重组蛋白,采用Westernblot和质谱分析确证该重组蛋白,采用MTT法检测所获得的重组蛋白对人脐静脉内皮细胞增殖的抑制活性。结果获得了相对分子质量为46000的重组人PEDF重组蛋白,该蛋白可显著抑制血管内皮细胞增殖。结论成功地经过原核表达获得了具有血管生成抑制活性的重组人色素上皮衍生因子。

Objective To express and purify the recombinant human pigment epithelium-derived factor(PEDF)which inhibits the proliferation of the endothelium cells from blood vessel in E.coli. Mothods PEDF gene was inserted into the prokaryotic expression vector pGEX-4T-2. The recombinant protein PEDF was expressed in E.coli BL-21, and purified by the GST Sepharose 4B affinity column. The recombinant human PEDF protein was identified by Western blot and mass spectrum. The biological activity of the recombinant human PEDF protein was measured by using MTT. Results The 46 kDa recombinant human PEDF protein was obtained. It significantly inhibited the proliferation of the human umbilical vein cell line HUVEC. Conclusion The recombinant human PEDF with anti-angiogenesis activity protein may be successfully purify through prokaryotic expression.