新型血管内皮细胞抑制因子VEGI_(151)的基因克隆表达及其活性

血管内皮细胞生长抑制因子 (vascularendothelialcellgrowthinhibitor,VEGI)是近年发现的一类肿瘤坏死因子超家族成员 ,具有抑制内皮细胞增殖的作用。从人脐静脉内皮细胞株 (ECV30 4)克隆到其基因 ,构建N端缺失 2 3个氨基酸的表达载体 ,并通过原核表达系统进行表达 (命名为VEGI151) ,表达量为 2 5 .5 % ,纯化后纯度达92 .5 %。通过生物学效应检测 ,发现VEGI151可明显抑制无血清培养基中内皮细胞的增殖 ,2 4h时VEGI151对内皮细胞的IC50 为 10mg/L ,0 .6 13mg/L时使内皮细胞在 36h内完全凋亡。通过检测体外培养肿瘤细胞 (A5 49、HepG2、Hela等 )的存活率 ,未发现明显的增殖或抑制效应。提示VEGI是一种主要作用于血管内皮细胞 ,在新生血管性疾病及肿瘤的治疗中有潜在的应用前景。

Cloning, Expression and Biological Activity of VEGI_(151), a Novel Vascular Endothelial Cell Growth Inhibitor

VEGI(vascular endothelial cell growth inhibitor) is a novel cytokine which belongs to the TNF super-family. In this study, the VEGI gene from ECV30 4 cells was cloned. A truncated form of VEGI, where 23 amino acids f rom N-terminal were deleted and named VEGI 151, was expressed in E.coli with 25.5% of expr ession rate. The purity of VEGI 151 reached 92.5% after purification. V EGI 151 sho wed significant inhibitory effect on endothelial cells. IC 50 of VEGI 15 1 was 10 m g/L at 24 h. At the concentration of 0.613 mg/L, VEGI 151 induced apopt osis of endothelial cells within 36 h. However, neither stimulatory effect nor i nhibitory effect of VEGI 151was detected on tumor cells(A549, HepG2, Hela)cultured in vitr o. These results suggested that endothelial cells was the main target cells of VE GI 151. Our findings indicate that VEGI 151 is a potential therapeutic drug on angiogenic disease and cancer.