人VEGF166基因转染兔骨髓内皮祖细胞及对其增殖的影响  

Trasfection of human VEGF166 gene into rabbit endothelial progenitor cells and its effects on cells proliferation

作　　者：曾庆娟[1] 尤捷[2] 许崇永[1] 方必东[1] 杨超颖[1] 杜玉清[3] 

机构地区：[1]温州医学院附属第二医院放射科,325027 [2]温州医学院附属第一医院肿瘤外科 [3]温州医学院附属第一医院介入科

出　　处：《浙江医学》2009年第7期927-929,共3页Zhejiang Medical Journal

基　　金：浙江省自然科学基金资助项目（Y205193）

摘　　要：目的探讨人血管内皮生长因子165（hVEGF165）基因转染兔骨髓源性内皮祖细胞（EPCs）及对EPCs增殖的影响。方法体外分离、培养、扩增并鉴定EPCs，脂质体介导转染携带增强型绿色荧光蛋白标记的VEGF166质粒、空白质粒，同时经ELISA法检测转基因EPCs表达VEGF蛋白，并采用CCK-8法检测对其增殖的影响。结果FITC标记剂豆凝集素I和Dil标记的乙酰化低密度脂蛋白荧光双染证实正在分化的EPCs，同时经免疫组化法证实VEGF受体2FLk-1和VIII因子的表达；VEGF165质粒转染组EPCs上清液中表达VEGF166,VEGF165转染EPCs的转染率约22．5％；hVEGF165基因转染促进EPCs增殖。结论hVEGF165基因可成功转染EPCs，且可促进EPCs的增殖，为基因治疗血管性疾病提供了动物实验基础。Objective To transfect human VEGF166 （hVEGF165） gene into rabbit endothelial progenitor cells （EPCs）, and to investigate its effect on EPCs proliferation. Methods EPCs were isolated from rabbit bone marrow and cultured in vitro. EPCs were transfected by lipofeetamine with green fluorescent protein-labeled VEGF165, or with blank plasmid. After transfection the expression of VEGF was detected by ELISA, and the effect on EPCs proliferation was measured by CCK8 method. Results The double FITC-UEA-land Dil-ac-LDL flouorescence staining displayed differentiated EPCs, and immunohistochemistry confirmed the expression of vascular endothelial growth factor receptor 2 and VIII factor. The expression of VEGF was detected in supernatant of hVEGF165-transfected EPCs with a expression rate of 22.5%. Conclusion hVEGF165 gene has been successfully transfected into EPCs, which enhances the proliferation of EPCs.

关 键 词：内皮祖细胞 血管内皮细胞生长因子 基因转染 

分 类 号：R733.3[医药卫生—肿瘤] R543.502[医药卫生—临床医学]