缺氧环境下A549细胞促进人脐静脉内皮细胞迁移及血管形成

目的 研究在缺氧条件下A549 细胞对人脐静脉内皮细胞（ HUVEC） 迁移及微血管形成的影响。方法 A549 细胞在常氧组（ 21% O2 ） 、缺氧组（ 2% O2 ） 、无氧组（ 0% O2 ） 中培养24 h 后, 通过倒置相差显微镜观察细胞形态、MTT 法检测细胞增殖、免疫细胞化学技术检测细胞内低氧诱导因子1α（ HIF-1α） 蛋白量以确定缺氧模型建立是否成功。随后取常氧组、缺氧组的A549 细胞上清液、HUVEC 细胞培养液干预HUVEC, 用细胞划痕实验、微丝绿色荧光染色方法观察各组HUVEC 的迁移情况及伪足形成, 体外HUVEC 三维培养技术观察血管形成情况。结果 与常氧组比较, 缺氧组A549细胞的生长状态好, 增殖明显; 无氧组A549 细胞生长状态差, 细胞数量减少; 缺氧组及无氧组A549细胞均有HIF-1α表达, 且缺氧组表达更加明显。与细胞培养液干预组比较, 缺氧上清液干预组和常氧上清液干预组A549 细胞的HUVEC 均有不同程度的迁移、伪足结构形成及血管管腔样结构形成,且前者较后者明显。结论 缺氧环境下A549 细胞生长状态良好, 增殖明显, 而无氧环境下则不利于A549 细胞的生长。缺氧环境下A549 细胞能促进HUVEC 的迁移、伪足形成及血管形成。

A549 Cells Promote HUVEC Migration and Angiogenesis under Hypoxic Conditions

Objective To observe the effects of A549 cells under hypoxicconditions on the migration of human umbilical vein endothelial cells （ HUVECs ） and microvascular formation.Methods After cultured for 24 h in normoxia condition（ 21% O2 ） , hypoxia condition （ 2% O2 ） , and anaerobic condition （ 0% O2 ） , respectively, morphology of A549 cells was observed with inverted phase contrast microscope, proliferation was detected by MTTassay, and intracellular hypoxia-inducible factor-1α（ HIF-1α）protein was detected by immunocyto-chemical technique, for determining whether the hypoxia model is successful. Then A549 cells’supernatant in the normoxic group, the hypoxia group and HUVECs culture medium were taken to intervene HUVECs. The migration of HUVECs was observed with cell scratch test,pseudopodia formation of HUVECs was observed with microfilament green fluorescent staining method, and blood vessel formation was observed with three-dimensional culture techniques in vitro.Results Compared with the normoxic group, the growth of A549 cells was better in the hypoxia group with more proliferation, and was poor in the anaerobic group with decreased number of cells. A549 cells in the hypoxia group and the anaerobic group both expressed HIF-1αprotein, which was more obvious in the anaerobic group. Compared with the HUVECs supernatant intervention group, the hypoxia supernatant intervention group and the normoxic supernatant intervention group both had varying degrees of migration, pseudopodia structure formation and vascular lumen sample structure formation , which were more obvious in the former group.Conclusions A549 cells in hypoxic environment grow very well, proliferated significantly, but anaerobic environment is not conducive to the growth of A549 cells which found to be apoptosis. A549 cells in hypoxic environment can promote HUVECs migration, pseudopodia formation and angiogenesis.