血管内皮生长因子121反义核酸抑制人胰腺癌细胞体外促血管生成能力

目的:探讨VEGF121反义核酸对体外培养胰腺癌细胞促血管生成能力的影响,以期通过抑制VEGF引起的肿瘤血管生成来达到抑制肿瘤生长的目的。方法:将稳定转染有VEGF反义核酸pcDNA3-ASVEGF121的胰腺癌细胞系BxPC-3在体外条件下培养,采用免疫组化检测肿瘤细胞表面VEGF及Flk-1表达水平。通过测定ECV-304生长曲线检测培养液上清对血管内皮细胞生长的影响。通过培养液上清体外血管生成试验检测肿瘤细胞体外促血管生成能力。结果:VEGF反义核酸转染BxPC-3细胞后,肿瘤细胞表面VEGF蛋白水平受到明显抑制,VEGF受体Flk-1表达水平上调。对血管内皮细胞ECV-304的促生长作用减弱。体外血管生成试验显示VEGF反义核酸能抑制肿瘤细胞体外促血管生成能力。结论:人反义VEGF121能显著抑制胰腺癌细胞表面的VEGF表达,抑制肿瘤细胞体外促血管生成能力。

Human Antisense VEGF121 Expression Vector Inhibits Angiogenensis Potential of Pancreatic Cancer Cell in vitro

Objective:To explore the effect of antisense VEGF_(121)on angiogenensis potential of pancreatic cancer cell in vitro,so as to depress the growth of tumor by inhibiting angiogenesis caused by VEGF.Methods:Pancreatic cancer cell BxPC-3 was transfected with pcDNA3-ASVEGF_(121)and pcDNA3 as control.VEGF and Flk-1 expression was tested by immunohistochemistry.Growth curve MTT assay of EVC-304 were carried out to assess the effect of culture media on vascular endothelial cell.In Vitro angiogenesis assay was performed to test angiogenensis potential of culture media.Results:Immunohistochemical stain revealed that antisense VEGF_(121)vector could inhibit VEGF expression in cell surface,and the expression of VEGF receptor,Flk-1,was found significantly elevated.ECV-304 growth curve revealed that proliferation effect of culture media on vascular endothelial cell was markedly reduced.In Vitro angiogenesis assay reveals that angiogenesis potential of culture media was significantly inhibited.Conclusion:Antisense VEGF_(121)vector can inhibit the VEGF expression of pancreatic cancer cell,and can also inhibit angiogenensis potential of pancreatic cancer cell in vitro.