中华医学杂志（英文版）2002，Vol.115Issue(6)：914-918,5.
VEGF逆转录病毒载体构建及在小鼠骨髓细胞中的表达
Expression of human VEGF121 cDNA in mouse bone marrow stromal cells
蔡善保 1马庆军 1余希杰 1党耕町 1马大龙2
作者信息
- 1. 北京大学第三医院骨科,北京,100083
- 2. 北京大学人类疾病基因中心,北京,100083
- 折叠
摘要
Abstract
Objective To construct a retroviral vector carrying human vascular endothelial growth factor (hVEGF121) cDNA for evaluation of the possibility of VEGF gene therapy in ischemic bone disease.Methods hVEGF121 cDNA was obtained from the plasmid pCDI/VEGF121 and cloned into retroviral plasmid pLXSN. Recombinant plasmid was transferred to the retrovirus packaging cell, PT-67, by lipofectamine mediated gene transfer. Mouse bone marrow stromal cells (MSCs) were transfected by the retrovirus. The integration of the hVEGF121 cDNA into MSC genomic DNA and expression of the VEGF gene was detected. Proliferation assays of human umbilical vein endothelial cells (HUVECs) by VEGF121 in culture medium were performed.Results Recombinant pLXSN/VEGF121 was correctly constructed and confirmed by restriction endonuclease analysis and DNA sequencing analysis. hVEGF121 gene was integrated into MSC genomic DNA after transfection, and the VEGF121 protein was expressed. Proliferation assays showed VEGF121 in culture medium was a biologically active protein and had a mitogenic effect on HUVEC.Conclusions Recombinant retroviral vector carrying hVEGF121 cDNA was successfully constructed. VEGF121 protein expressed by MSCs had mitogenic effect biologically. This provides a further foundation for VEGF gene therapy for bone ischemic disease and bone tissue engineering.关键词
内皮细胞生长因子/逆转录病毒/基因治疗/骨髓基质细胞Key words
endothelial growth factor/retrovirus vector/gene therapy/marrow stromal cell分类
医药卫生引用本文复制引用
蔡善保,马庆军,余希杰,党耕町,马大龙..VEGF逆转录病毒载体构建及在小鼠骨髓细胞中的表达[J].中华医学杂志（英文版）,2002,115(6):914-918,5.
