低氧对人肺腺癌A549细胞迁移和黏附的影响

Effect of hypoxia on migration and adhesion of human pulmonary adenocarcinoma A549 cells

目的:探讨低氧对人肺腺癌A549细胞迁移以及与血管内皮细胞黏附的影响。方法:细胞划痕实验和Transwell实验检测低氧对A549细胞迁移和浸润的影响;细胞黏附实验检测低氧对A549细胞与血管内皮细胞黏附的影响;免疫荧光法观察低氧对A549细胞中E-cadherin、β-catenin、纤维状肌动蛋白(F-actin)分布的影响;荧光素酶报告基因检测低氧对A549细胞中依赖于缺氧诱导因子-1(hypoxia-inducible factor-1,HIF-1)的转录活性的影响。结果:低氧可以促进A549细胞的迁移、浸润和与血管内皮细胞的黏附,影响A549细胞中E-cadherin和β-catenin的分布及F-actin细胞骨架的重排,上调A549细胞中依赖于HIF-1的报告基因的表达。结论:低氧促进A549细胞的迁移、浸润和与血管内皮细胞的黏附可能是通过调控A549细胞中依赖于HIF-1基因的表达,进而影响细胞中E-cadherin和β-catenin分布和F-actin细胞骨架的重排。

Objective： To evaluate the effect of hypoxia on migration, invasion and adhesion to endothelial cells of human pulmonary adenocarcinoma A549 cells. Methods： Wound-healing and Transwell invasion assays were performed to study the effect of hypoxia on migration and invasion of A549 cells. Cell adhesion test was used to detect the adhesion of A549 cells to a monolayer of human umbilical vein endothelial cells （HUVECs）. Immunofluorescence assay was used to evaluate the effect of hypoxia on distribution of E- cadherin, β-catenin, and F-actin. The luciferase reporter gene assay was performed to detect the transcription of hypoxia-inducible factor-1 （ HIF-1 ） alpha. Results： Hypoxia promotes A549 cell migration, invasion, and adhesion to endothelial cells, and modulated the distribution of E-cadherin and β-catenin and rearrangement of actin cytoskeletal protein, and up-regulated HIF-l-dependent reporter gene expression in A549 cells. Conclusion： Hypoxia promoted A549 cell migration, invasion, and adhesion to endothelial cells by upregulating HIF-1-dependent gene expression, subsequently affecting the redistribution of E-cadherin and β-catenin and rearrangement of F-actin cytoskeletal protein.