不同缺氧状态对人肝癌HepG2细胞侵袭和转移能力的影响

Effects of hypoxia in different degrees on in vitro invasion and migration of human hepatocarcinoma HepG2 cells

目的:探讨不同缺氧状态对人肝癌HepG2细胞黏附、侵袭和转移能力的影响及其可能机制。方法:不同浓度的低氧处理对数生长期的人肝癌HepG2细胞,采用MTT法、Transwell膜侵袭系统、免疫细胞化学方法、明胶酶谱分析和反转录PCR检测变异型白细胞分化抗原CD44v6、基质金属蛋白酶2(MMP-2)、MMP-9、低氧诱导因子(HIF-1α)和血管内皮生长因子(VEGF)的表达差异。结果:HepG2细胞经低氧处理后,细胞的基质黏附率、穿透基底膜与游走迁移的细胞数均增高,以3%氧浓度最为显著(P<0.05);3%和5%氧处理还可显著促进CD44v6的表达,增加MMP-2和MMP-9的表达,并可上调HIF-1α和VEGF。结论:适度缺氧能增强人肝癌细胞的黏附、侵袭和转移能力,其机制可能与CD44v6、基质金属蛋白酶、HIF-1α和VEGF的表达改变有关。

AIM： To study the effects and the underlying mechanisms of hypoxia on adhesion,invasion and migration of HepG2 human liver cancer cells cultured in vitro. METHODS： HepG2 cells were cultured under different concentrations of oxygen for 24 h.MTT assay was used to measure the cell adhesion rate.The Transwell chambers were used to assess in vitro invasion and migration.The expression of CD44v6,matrix metalloproteinase 2（MMP-2）,MMP-9,hypoxia-inducible factor-1α（HIF-1α） and vascular endothelial growth factor（VEGF） was detected by the methods of immunocytochemistry,gelatin zymography and RT-PCR. RESULTS： Hypoxia,especially under 3% oxygen,significantly enhanced the adhesion rates of HepG2 cells,and increased the number of the cells migrated through Matrigel-coated chambers（P0.05）.HepG2 cells treated with 3% and 5% oxygen increased the expression of CD44v6,MMP-9,MMP-2,HIF-1α and VEGF. CONCLUSION： Pretreatment with moderate hypoxia enhances in vitro adhesion,invasion and migration of HepG2 cells.Multiple molecules including CD44v6,MMP-2,MMP-9,HIF-1α and VEGF may be involved in the processes.