【摘要】 目的应用聚乙烯亚胺(polyethyleneimine,PEI)包裹淋巴管内皮祖细胞(lymphatic endothelial progenitor cells,LEPCs)血管内皮生长因子受体3(vascular endothelial growth factor receptor-3,VEGFR-3)基因siRNA,体外沉默VEGFR-3基因表达,研究抑制LEPCs分化抗肿瘤淋巴管新生的作用。方法 Ficoll法分离人脐血单个核细胞,流式细胞仪、免疫荧光分选并鉴定LEPCs。PEI包裹VEGFR-3siRNA并转染入LEPCs,流式细胞仪检测转染效率。RT-PCR和Western blot检测mRNA和蛋白质水平VEGFR-3基因沉默效果。结果 VEGFR-3siRNA成功转入LEPCs,转染效率30%。VEGFR-3siRNA转染组LEPCs VEGFR-3mRNA和蛋白表达均降低(P<0.05)。结论体外经由PEI介导的siRNA能有效的抑制LEPCs VEGFR-3的表达,为以LEPCs为靶点抑制肿瘤淋巴管新生提供了实验依据。更多还原

【Abstract】 Objective To study the gene silencing efficiency of siRNA targeted against human VEGFR-3 of lymphatic endothelial progenitor cells(LEPCs) in vitro mediated by polyethyleneimine(PEI),and to approach AntiLymphangiogenesis in Tumor. Methods Mononuclear cells were isolated from umbilical cord blood using density centrifugation with Ficoll solution. LEPCs were sorted and viewed using flow cytometer and confocal laser scanning microscope,respectively.VEGFR-3 siRNA/PEI were transfected LEPCs. Flow cytometry for transfection efficiency. RT-PCR and Western blot were used to evaluate VEGFR-3 gene silencing induced by VEGFR-3 siRNA. Results VEGFR-3 siRNA/PEI were transfected to LEPCs, the transfection efficiency was 30%. The expression of VEGFR-3mRNA and protein in siRNA/PEI-transfected LEPCs were significantly down-regulated compared with control groups(P < 0.05). Conclusion The PEI could effectively transfect siRNA into cells and silence the VEGFR-3 gene expression of LEPCs. The research provided experimental evidence for clinical study of tumor lymphangiogenesis about LEPCs.更多还原