摘要
目的构建血管内皮生长因子(vascu lar endothelial growth factor,VEGF)真核表达载体,并研究其在骨髓间充质干细胞(m esenchym al stem cells,MSCs)中的表达情况。方法运用DNA重组技术,构建VEGF真核表达载体pcDNA3.1(-)/hVEGF165,将载体转染大鼠MSCs,RT-PCR,ELISA及免疫细胞化学等方法检测VEGF在大鼠MSCs中的表达情况,MTT法检测转染后细胞上清液中VEGF的生物活性。结果成功构建VEGF真核表达载体pcDNA3.1(-)/hVEGF165,其转染后的大鼠MSCs中VEGF表达水平明显增高,转染后细胞培养上清液具有促使内皮细胞增殖的生物活性。结论VEGF真核表达载体转染大鼠MSCs后能有效增加VEGF表达,并表现出较强的促内皮细胞增殖作用。
AIM To construct eukaryotic expression vector of （VEGF） gene and to investigate its transfection and expression METHODS After pcDNA3.1 （ - ）/hVEGF165 vector was human vascular endothelial growth factor in rat mesenchymal stem cells （MSCs）. successfully constructed, rat MSCs were transfected with jet-PEI. RT-PCR, ELISA and immuocytochemical methods were used to detect the expression of VEGF and MTT was used to detect the biological activity. RESULTS VEGF mRNA and protein in the transfected group significantly increased and obvious biological activity was observed in stimulating proliferation of endothelial cells. CONCLUSION pcDNA3.1 （ - ）/hVEGF165 vector can effectively transfect rat MSCs. The transfected rat MSCs significantly increase VEGF mRNA and protein expression, and have relatively strong biological activity.
出处
《心脏杂志》
CAS
2006年第2期135-137,共3页
Chinese Heart Journal
