低氧应激对人肝癌细胞AFP、VEGF、TIMP-1、MMP-9表达的影响

Effects of hypoxia stress on expressions of AFP,VEGF,TIMP-1 and MMP-9 in HepG-2 cell

目的:探讨低氧应激对人肝癌细胞(HepG-2细胞)甲胎蛋白(AFP)、血管内皮生长因子(VEGF)、金属蛋白酶组织抑制剂(TIMP)-1、基质金属蛋白酶(MMP)-9表达的影响。方法:低氧(5%O2、5%CO2、90%N2)和常氧培养HepG-2细胞。逆转录-多聚酶链式反应技术测定HepG-2细胞AFP、VEGF、TIMP-1、MMP-9 mRNA的表达。酶联免疫吸附法测定HepG-2细胞培养上清VEGF、TIMP-1、MMP-9蛋白含量,放射免疫测定法测定HepG-2细胞培养上清AFP蛋白含量。结果:低氧组AFP、VEGF、MMP-9 mRNA和蛋白表达量显著高于常氧组;TIMP-1 mRNA和蛋白表达量显著低于常氧组(P<0.01)。结论:低氧应激上调人HepG-2细胞AFP、VEGF、MMP-9基因表达,下调TIMP-1基因表达。

Objective：To investigate whether expressions of alpha-fetoprotein（AFP）,vascular endothelial growth factor（VEGF）,tissue inhibitor of metalloproteinase（TIMP）-1 and matrix metalloproteinase（MMP）-9 in HepG-2 cell are regulated by hypoxia stress or not.Methods： HepG-2 cells were cultured under hypoxia（5% O2,5% CO2 and 90% N2） or normoxia.The expressions of AFP,VEGF,TIMP-1 and MMP-9 mRNA in HepG-2 cell were detected with reverse transcription polymerase chain reaction（RT-PCR）.The expressions of VEGF,TIMP-1,MMP-9 protein in supernatant of HepG-2 cell were studied with enzyme-labeled immunosorbent assay（ELISA）.The expression of AFP protein in supernatant of HepG-2 cell was determined with radioimmunity assay（RIA）.Results： The expressions of AFP,VEGF and MMP-9 in hypoxia group were dramatically higher than those in normoxia group,while the expression of TIMP-1 in hypoxia group was significantly lower than that in normoxia group（P0.01）.Conclusion： The expressions of AFP,VEGF and MMP-9 gene in HepG-2 cell are up-regulated while that of TIMP-1 is down-regulated by hypoxia stress.