Expression And Purification Of HVEGF₁₂₁/EGFP Fusion Protein And Researching Its Function

OBJECTIVE: To transfect bone marrow-derived mesenchymal stem cells (BMCS) with recombinant plasmid containing human vascular endothelial cell growth factor121 (hVEGF121) gene and enhanced green fluorescence protein report (pEGFP) gene, to detect the expression of the fusion protein hVEGF121/EGFP, and to study its function on cell proliferation and vascular permeability in vitro.METHODS: Recombinant plasmid of pEGFP/hVEGF121 built preliminary was amplified, and BMSCs were transfected with the plasmid by positive ionic liposome transfection method. The expression of fusion protein in the cells was detected by fluorescence microscope and Western blot. The function of the protein on cell proliferation and vascular permeability was determined with MTT and Miles test respectively.RESULTS: Fluorescence microscopy showed that the expression of green fluorescence in the cells transfected with pEGFP-N2-hVEGF121, and Westen blot confirmed the expression of VEGF in the culture media. The results of MTT exhibited that the fusion protein stimulated cell proliferation more effectively compared with the control protein (P < 0.05). Miles test suggested that the fusion protein could increase the permeability of the blood vessel wall.CONCLUSION: Fusion protein hVEGF121/EGFP was successfully expressed in BMSCs. Furthermore, the protein kept the function of wild-type VEGF in vitro. Thus, the BMSCs transfected with hVEGF121/EGFP could be very useful in future studies of cardiovascular diseases.