bFGF基因转染对血管内皮细胞迁移的影响及机制  

Effects of bFGF gene transfection on the migration of vascular endothelial cells and its mechanism

作　　者：徐彬[1] 武晓英[2] 林桂先[2] 毛建文[2] 

机构地区：[1]广东药学院生命科学与生物制药学院,广州510006 [2]广东药学院基础学院

出　　处：《山东医药》2010年第11期21-24,共4页Shandong Medical Journal

基　　金：广东药学院博士启动基金资助项目(2006SMK03)

摘　　要：目的研究碱性成纤维细胞生长因子(bFGF)基因转染对人脐静脉内皮细胞(HUVECs)迁移能力的影响,并探讨其可能机制。方法通过基因亚克隆构建真核表达载体pcDNA3.1-bFGF,利用脂质体介导将bFGF基因导入HUVECs内,通过RT-PCR和ELISA法检测基因的表达;bFGF基因转染后的HUVECs通过Transwell小室法检测其迁移能力的变化;W estern blot法检测转染后的细胞Raf-1、细胞外调节蛋白激酶2(ERK2)和黏着斑激酶(FAK)蛋白表达变化。结果成功构建了表达载体pcDNA3.1-bFGF,该载体转染HUVECs后,bFGF mRNA和蛋白均显著增加。转染bFGF基因可使HUVECs的体外迁移能力增强,上调ERK2和FAK蛋白的表达,但对Raf-1蛋白表达无影响。结论bFGF基因转染能促进血管内皮细胞的迁移,其作用机制可能与上调ERK2和FAK蛋白表达有关。Objective To investigate the effects and its posssible mechanisms of basic fibroblast growth factor（ bFGF） gene transfeetion on the migratory activity of vascular endothelial cells （HUVECs）. Methods Eukaryotie expression plasmid peDNA3.1-bFGF was constructed by gene subclone, and then was transfected into HUVECs by liposome mediating, RT-PCR and ELISA were used to determine the expression of bFGF mRNA. Transwell Chamber assay was performed to detect the cell migratory capacity changes of HUVECs after bFGF gene transferred. The expression level of Raf-1, extracellular regulated protein kinases 2 （ERK2） and focal adhesion kinase （FAK） protein were assessed by Western blot. Results pcDNA3.1-bFGF was successfully constructed, bFGF mRNA and protein both increased significantly/ffter the transfection. bFGF gene transfection promoted the migratory capacity of HUVECs in vitro, and up-regulated the expression of ERK2 and FAK protein ERK2 and FAK protein, but not affected the expression of Raf-1 protein. Conclusions The transfection of bFGF gene can promote the migration of vascular endothelial cell, its mechanisms might be associated with up- regulation of the expression of ERK2 and FAK protein.

关 键 词：碱性成纤维细胞生长因子 基因转染 血管内皮细胞 迁移 

分 类 号：B322.1[哲学宗教—外国哲学]