缺氧对肺癌A549细胞外泌体的影响  

Effect of hypoxia on the extracellular vesicles of lung cancer A549 cells

作　　者：张乐 郑亚妮 王荣华 王耿杰 洪环谅 马良赟 Zhang Le;Zheng Yani;Wang Ronghua;Wang Gengjie;Hong Huanliang;Ma Liangyun(Department of Thoracic Surgery,the 910th Hospital of the Chinese People's Liberation Army Joint Logistic Support Force,Quanzhou 362000;Department of Human Anatomy,Histology and Embryology,Tongji University,Shanghai 200092,China.)

机构地区：[1]联勤保障部队第九一〇医院心胸外科,泉州362000 [2]同济大学人体解剖学与组织胚胎学教研室,上海200092

出　　处：《解剖学杂志》2024年第3期231-234,共4页Chinese Journal of Anatomy

基　　金：2024年泉州市科技计划项目(2024NY03)。

摘　　要：目的:探讨缺氧对肺癌A549细胞外泌体形态、表型及蛋白表达的影响。方法:对A549细胞进行缺氧培养(实验组)和常氧培养(对照组)后,分别用外泌体提取试剂盒提取外泌体,然后用透射电镜观察其结构形态变化,免疫印迹检测其标记蛋白表达变化,BCA蛋白定量法检测外泌体蛋白量的变化。结果:实验组和对照组A549细胞提取的外泌体直径皆为30~120 nm,呈圆形或椭圆形,有完整胞膜结构,形态规则,内含低密度物质,边缘比较清晰;实验组中A549外泌体的CD9蛋白表达与对照组无明显差异,但是CD81的蛋白表达明显高于对照组。BCA检测结果显示,A549细胞分泌的外泌体蛋白量与接种细胞数呈线性关系,在缺氧状态下A549细胞分泌外泌体的蛋白量明显高于对照组。结论:缺氧导致肺癌细胞A549外泌体CD81的表达增多,细胞分泌外泌体蛋白量增多,提示缺氧状态下的A549细胞具有更强的远处转移能力。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 A549lung cancer cells,and the amount of exosomal protein secretion increases,suggesting that A549 cells under hypoxic conditions have a stronger ability for distant metastasis.

关 键 词：A549细胞 外泌体 缺氧 蛋白表达 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]