Under Hypoxic Conditions A549cells Promote HUVEC Migration And Angiogenesis

Objective:The aim of this study is to observe that A549cells under hypoxicconditions promote human umbilical vein endothelial cells migration, pseudopod formation and vascular formation through establishing hypoxia model of the A549cells in vitro, which reveals the relationship during hyp-oxia,endothelial cell migration, pseudopod formation and vascular formation,and is an important part of angiogenesis.Methods:After A549cells cultured for24hours at different oxygen concentrations (21%,2%,0%) conditions, morphology of A549cells were observed by inverted phase contrast microscope, proliferation were detected by MTT assay, and intracellular HIF-1a protein were detected by immunocytochemical techniques,which can prove the success of hypoxia model. Then the supernatan of A549cell in the normoxic and hypoxic,group was collected,and was taken to intervened HUVEC at different concentrations (50ul/ml,100ul/ml,150ul/ml),but in control group cell culture medium instead. Comparing migration distance of HUVEC in each group by cell scratch test method, which can find the optimal concentrationof supernatan in each group.Hereafter taking this optimal concentration of supernatan deal with HUVECs,observing its pseudopod formation by the microfilament green staining method, counting and comparing vascular formation in each group under inverted phasecontrast microscope by three-dimensional culture techniques in vitro.Results:At the experimental parts of hypoxia model establishment:1) compared with the normoxic group, A549cells in hypoxia group growed very well and increased,some of them had irregular shape named pseudopod, showing that hypoxia is conducive to A549cell invasion and metastasis),but in anaerobic group, A549cells decreased significantly and shrinkaged, their morphology partly disappeared and nuclear chromatin condensationed, in addtion some of them floated through culture medium.2) compared with the normoxic group, the OD values in hypoxia group was higher (P<0.05),but significantly lower in the anaerobic group (P<0.05);3) compared with the normoxic group, the expression of HIF-1a protein in the hypoxia and anaerobic group were well obvious (P <0.05),but in hypoxia group is the mose obvious (P<0.05).At the experimental parts of HUCVE migration and angiogenesis:1)by the comparison between groups and within the group,in the control group, HUVEC migration was not obvious and was no significant difference at different concentration(P>0.05), while HUVEC migration was obvious and concentration-dependent manner in the hypoxia supernatant intervention group and in the normoxic supernatant intervention group,the former is more significant than the latter (P<0.05);2) compared with the control group, HUVEC were seen pseudopod formation in the hypoxia supernatant intervention group and in the normoxic supernatant intervention group, and the former was more than the latter.3) With HUVEC three-dimensionalculture techniques in vitro, compared with the control group, the lumen-like vascular structures per field under inverted phase contrast microscope was found in other two groups,and the hypoxia supernatant intervention group was increased more than the normoxic supernatant intervention group(P <0.05).Conclusion:1In hypoxic environment A549cells grow very well, significantly proliferate,but anaerobic environment is not conducive to the growth of A549cells which found to be apoptosis.2A549cells growing in hypoxic environment. can promote HUVEC migration, pseudopodia formation, angiogenesis in hypoxic environment.