缺氧应激对人肝癌细胞黏附、侵袭和迁移能力的影响

Effects of hypoxia stress on adhesion,invasion and migration of human hepatocellular carcinoma cell HepG2

目的探讨缺氧应激对人肝癌细胞HepG2黏附、侵袭和迁移能力的影响。方法缺氧(5%O2、5%CO2、90%N2)和常氧培养HepG2细胞。细胞-基质黏附实验测定常氧组和缺氧4、12、24h组HepG2细胞黏附能力;细胞侵袭和迁移实验测定缺氧和常氧24 h组HepG2细胞侵袭和迁移能力;逆转录聚合酶链反应(RT-PCR)测定常氧组和缺氧4、12、24 h组HepG2细胞缺氧诱导因子(hypoxia inducible factor,HIF)-1α mRNA的表达量。结果随着缺氧时间的延长HepG2细胞的黏附能力明显增强(P<0.01),缺氧24 h组HepG2细胞侵袭和迁移能力明显高于常氧组(P<0.01)。常氧下HepG2细胞不表达HIF-1α mRNA,缺氧4 h时HIF-1α mRNA表达量最高,随后下降(P<0.01),但仍高于常氧组。结论缺氧应激上调HepG2细胞HIF-1α mRNA的表达,促进HepG2细胞黏附、侵袭和迁移。

Objective To investigate whether hypoxia stress enhances adhesion, invasion and migration of human hepatocellular carcinoma cell HepG2 or not. Methods HepG2 cells were cultured under hypoxia （5% 02, 5% CO2 and 90% N2） for 4, 12, 24 h or normoxia. The adhesion of HepG2 cells was studied by cell-matrix adhesion assay. Cell invasion and migration assay was used to determine the changes in invasion and migration of HepG2 cells respectively. The expression of HIF-1 α mRNA in HepG2 cells was determined by reverse transcription polymerase chain reaction （RT-PCR）. Results The adhesion of HepG2 cells was gradually enhanced with the increasing hypoxia time （P 〈0. 01 ）. The invasion and migration of HepG2 cells after 24-hour hypoxia was significantly higher than that after 24-hour normoxia （ P 〈 0. 01 ）. The expression of HIF-1 α mRNA reached the highest level when the cells were exposed to hypoxia for 4 h, then decreased, but still higher than that in normoxia （P 〈 0. 01 ）. Conclusion Hypoxia stress upregulates the expression of HIF-1 α mRNA, and enhances the adhesion, invasion and migration of HepG2 cells.