TGF-β1和低氧对肝癌HepG2细胞表达VEGF的影响

Effects of transforming growth factor 1 and hypoxia on the expression of vascular endothelial growth factor for Hepatoma cells

目的探讨转化生长因子β1(TGF-β1)和低氧(CoCl2)对肝癌细胞血管内皮生长因子(VEGF)表达的影响。方法体外培养肝癌细胞系HepG2细胞,实验分为对照组、不同浓度TGF-β1和CoCl2的分别处理组、TGF-β1(100 pmol/L)+CoCl2(200 μmol/L)组;用免疫细胞化学法检测VEGF蛋白表达;半定量RT-PCR技术检测细胞中VEGFmRNA相对表达量。结果各处理组VEGF蛋白表达均为阳性;TGF-β1或CoCl2组中VEGF mRNA相对表达量明显高于对照组(P<0.01),并且呈浓度依赖性;TGF-β1+CoCl2组中VEGFmRNA表达高于单因素刺激组(P<0.05)。结论TGF-β1能够刺激HepG2细胞表达VEGF,并且可以协同CoCl2增加VEGF的表达。

Objective To observe the effect of Vascular Endothelial Growth Factor （VEGF） to the expression in hepatoma cells affected by TGF-β1 and hopxia. Methods HepG2 cells were cultured in vitro, and treated with different doses of TGF-β1 and Cobalt chloride hexahydrate（ CoCl2 ）, or with TGF-β1 and COCl2. The change of VEGF protein and mRNA was detected by immunohistochemistry and semiquantitative reverse transcriptase-polymerase chain reaction （RT-PCR）. Results Immunohistochemistry and RT-PCR showed that the expression of VEGF protein and mRNA were significantly higher in TGF-β1 groups or CoCl2 groups than that of control group（ P 〈0. 01 ）, they increased the expression of VEGF in concentrate-dependently manner. The incubation with TGF-β1 and CoCl2 results in significantly more VEGF expression than either stimulation alone （P 〈 0. 05 ）. Conclusion TGF-β1 can increase the expression of VEGF mRNA by Hepatoma cells; TGF-β1 can promote hypoxia induced by CoCl2to increase the expression of VEGFmRNA.