Effect of hypoxia on the extracellular vesicles of lung cancer A549 cells
Objective:To investigate the impact of hypoxia on the morphology,phenotype,and protein expression of extracellular vesicles(EVs)secreted by A549 lung cancer cells.Methods:After culturing A549 cells under both hypoxic(experimental group)and normoxic(control group)conditions,extracellular vesicles were isolated using an exosome extraction kit.The structural morphological changes were observed via transmission electron microscopy,while alterations in marker protein expressions were detected through immunoblotting.The protein content of the extracellular vesicles was quantified using the bicinchoninic acid(BCA)assay.Results:EVs isolated from both experimental and control groups demonstrated diameters within the range of 30-120 nm,exhibiting rounded or elliptical shapes with intact membrane structures,consistent morphology,low-density interiors,and distinct boundaries.No significant change was observed in the expression of the CD9 protein between the hypoxic and normoxic groups.However,a marked increase in CD81 protein expression was noted in the hypoxic group compared with the control groups.The BCA assay revealed that there was a linear correlation with cell seeding density,and the protein level of exosomes secreted by A549 cells was significantly higher than that of the control group under hypoxic conditions.Conclusion:Hypoxia increases the expression of CD81 in exosomes from A549 lung cancer cells,and the amount of exosomal protein secretion increases,suggesting that A549 cells under hypoxic conditions have a stronger ability for distant metastasis.
万方数据
维普
