关键词: 生长因子, 内皮, 血管, 转染, 骨髓基质细胞, K562细胞

Abstract: Objective To get the protein expression of sFlt-1 by transfection and to investigate the effects of sFlt-1 gene transfection on the growth of K562 cells. Methods The recombinant plasmid pcDNA₃-sFlt-1_D4 was constructed.The recombinant plasmid pcDNA3-sFlt-1_D4 was transfected into the bone marrow stromal cells by Lipofectamine 2000，which was identified by RT-PCR,ELISA and MTT.Results The transfection efficiency identified by flow cytometry was 9.27%.The protein expression of sFlt-1D4 was found in the culture supernatant 24 h,48 h and 4 weeks after transfetion by ELISA and the expression concentrations were（0.104±0.078 ），(0.158±0.022) and（0.171±0.069） μg•L^-1，respectively.The content of VEGF secreted by K562 culturing with transfectant cells culture supernatant was reduced compared with control.The inhibitoy rates on the proliferation of K562 cells via MTT assay were 9.41%±4.71%，23.63%±7.50%， and 33.13%±6.93%，respectively. Conclusion The bone marrow stromal cells transfected with recombinant plasmid pcDNA3-sFlt-1_D4 could secrete sFlt-1_D4 and inhibit the proliferation of K562 cells.

Key words: growth factor, endothelium, vascular, transfection, bone marrow stromal cells, K562 cells