hVEGF165/pcDNA 3.1在小鼠心肌细胞中的表达

hVEGF165/pcDNA3.1 Expression in Neonate Mice Myocardial Cells

目的观察重组质粒人血管内皮生长因子165基因与质粒载体pcDNA 3.1的重组体(hVEGF 165/pcDNA 3.1)在心肌细胞中的表达,为冠心病的基因治疗和细胞移植治疗研究奠定基础。方法体外培养小鼠心肌细胞,脂质体介导hVEGF 165/pcDNA 3.1转染心肌细胞,用逆转录-聚合酶链反应(RT-PCR)、细胞免疫化学方法和免疫印迹法(W estern b lotting)检测其在心肌细胞中的瞬时表达。结果RT-PCR扩增得到与hVEGF 165大小一致的目的条带,细胞免疫化学法检测到心肌细胞胞质中hVEGF蛋白,W estern b lotting在培养的细胞上清液中检测到hVEGF蛋白。结论构建的重组质粒hVEGF 165/pcDNA 3.1能在体外培养的小鼠心肌细胞中表达,可用于冠心病的基因和细胞移植治疗研究。

Objective To investigate the transfection and expression of recombinant plasmid human vascular endothelial growth factor 165/pcDNA3. 1 （hVEGF165/pcDNA3. 1） in myocardial cells, and to build foundation for gene therapy and cell therapy of coronary artery disease （CAD）. Methods Myocardial cells were cultured in vitro and transfected by hVEGF165/pcDNA3.1 with liposome; then transient expressed protein was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）, immunochemistry and Western blotting. Results A strap as hVEGF165 was obtained by RT-PCR, the protein of hVEGF165 was found in myocardial cells by immunochemistry and in supernatant by Western blotting. Conclusion The recombinant plasmid hVEGFI65/pcDNA3. 1 can be expressed in myocardial cells, and may be used in studying CAD by gene therapy and cell transplantation.