中国免疫学杂志2011，Vol.27Issue(8)：686-690,5.DOI:10.3969/j.issn.1000-484X.2011.08.003
肝细胞生长因子基因转染对人脐静脉内皮细胞增殖、迁移的影响
Hepatocyte growth factor (HGF) gene promoted proliferation and migration in human umbilical vein endothelial cells using plasmid mediated transfection
摘要
Abstract
Objective:To investigate the effects of hepatocyte growth factor(HGF)gene on the proliferation and migration of human aumbilicalvein endothelial cells(ECV 304).Methods:The plasmid pRc/CMV-HGF was used as the template to amplify HGF gene by PCR.The products were cloned in eukaryotic expression vector pEGFP-N1 to construct recombinant plasmids as pEGFP-HGF.The recombinaint plasmids pEGFP-HGF were identified by restrictive endonuclease digestion and sequencing,then transfected into human umbilical vein endothelial cellswith liposome,and positive clones were selectited by G418 .The transfection and the expression of HGF IN ECV 304 were tested by PT-PCR and immunohistochemistry.ELISA was performed to detected levels of HGF in ECV 304.Proliferative activites of the cells were evaluated by MTT assay.Transwellmigration assay was performed to detectmigration activity.Results:The recombinaintplasmids were qualified by restrictive endonuclease digestion and squencing.After G418 selection,the cell clones were obtained succesfully.Under a fluorescent microscope,the expression of enhanced green fluorescent protein(EGFP)was seen in cells transfected with vector contain EGFP gene.RT-PCR and immunohistochemistry proved that there was transcription of HGF gene in transfection cells and there was expression of HGF in these cells.The HGF levels in the HGF/ECV 304 cells increased significantly at the 48 th hour after thansfection.MTT and transwellmigration assay showed HGF promoteproliferation and migration of the cultured human umbilicalvein endothelial cells.Conclusion:The recombinant of pEGFP-HGF is an effective expression vector.HGF produced by the transfected cells promotes the proliferation and migration of the human umbilicalvein endothelial cells,HGF is thus a potent agent for strategies designed to promote therapeutic angiogenesis.关键词
HGF基因/内皮细胞/增殖/迁移Key words
Hepatocyte growth factor(HGF)/Endothelial cells/Proliferation/Migration分类
医药卫生引用本文复制引用
常冰梅,李美宁,刘志贞,张悦红,程牛亮,牛勃..肝细胞生长因子基因转染对人脐静脉内皮细胞增殖、迁移的影响[J].中国免疫学杂志,2011,27(8):686-690,5.基金项目
本文由细胞生理学省部共建教育部重点实验室主任基金(2010-01)、山西省科技攻关项目(20080311059-6)和山西医科大学科技创新项目(01200713)资助 （2010-01）
