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Title:: Construction of VEGF eukaryotic expression vector and its expression in mesenchymal stem cells

作者:: 高峰; 王红兵; 俞世强; 蔡振杰; 第四军医大学西京医院心血管外科研究所，陕西西安 710032

Author(s):: GAO Feng; WANG Hong-bing; YU Shi-qiang; CAI Zhen-jie; Institute of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi′an, Shaanxi 710032, China

关键词:: 血管内皮细胞生长因子; 真核表达载体; 骨髓间充质干细胞

Keywords:: vascular endothelial growth factor; eukaryotic expression vector; mesenchymal stem cells

分类号:: Q58

DOI:: -

文献标识码:: A

摘要:: 目的构建血管内皮生长因子（vascular endothelial growth factor，VEGF）真核表达载体，并研究其在骨髓间充质干细胞（mesenchymal 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。

Abstract:: AIM To construct eukaryotic expression vector of human vascular endothelial growth factor (VEGF) gene and to investigate its transfection and expression in rat mesenchymal stem cells (MSCs). METHODS After pcDNA3.1(-)/hVEGF165 vector was 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.

参考文献/References

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［2］ Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells［J］. Science, 1999, 284:143-147.

［3］ Wollert KC, Meyer GP, Lotz J, et al. Intracoronary autologous bonemarrow cell transfer after myocardial infarction: the BOOST randomised controlled clinical trial［J］. Lancet, 2004, 364 (9429):141-148.

［4］ Neufeld G, Cohen T, GitayGoren H,et al. Similarities and difference between the ascular endothelial growth factor(VEGF) splice variants ［J］. Cancer Metastasis Rev,1996,15(2): 153-158.

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VEGF真核表达载体构建及其在骨髓间充质干细胞中的表达(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

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