摘要
目的探讨鸢尾素对早期B细胞因子3(EBF3)/花生四烯酸-15-脂加氧酶(ALOX15)通路的调控作用及肺腺癌细胞增殖和迁移的影响。方法将A549细胞分为鸢尾素溶剂组、鸢尾素组、sh-NC组、EBF3抑制剂(sh-EBF3)组、鸢尾素+sh-NC组、鸢尾素+sh-EBF3组。5-溴-2-脱氧尿嘧啶(EdU)染色、CCK-8法检测细胞增殖;划痕实验检测细胞划痕愈合率;2′,7′-二氯荧光二乙酸酯(DCFH-DA)染色检测细胞中活性氧(ROS)水平;比色法检测细胞中谷胱甘肽(GSH)、丙二醛(MDA)、细胞亚铁(Fe^(2+))水平;透射电镜观察A549细胞中线粒体形态;qRT-PCR检测A549细胞中增殖性细胞核抗原(PCNA)、基质金属蛋白酶2(MMP-2)、谷胱甘肽过氧化酶4(GPX4)mRNA水平;Western blot检测细胞中EBF3、ALOX15蛋白。结果与鸢尾素溶剂组比较,鸢尾素组A549细胞线粒体呈现出铁死亡特征,EdU阳性率、OD_(450)值、划痕愈合率、GSH水平、PCNA、MMP-2、GPX4 mRNA水平降低,ROS相对荧光强度、MDA、Fe^(2+)水平及EBF3、ALOX15蛋白升高(P<0.05);与sh-NC组比较,sh-EBF3组A549细胞线粒体呈现出铁死亡现象有所减弱,EdU阳性率、OD_(450)值、划痕愈合率、GSH水平、PCNA、MMP-2、GPX4 mRNA水平升高,ROS相对荧光强度、MDA、Fe^(2+)水平及EBF3、ALOX15蛋白降低(P<0.05);sh-EBF3逆转了鸢尾素对A549细胞铁死亡、增殖与迁移的影响。结论鸢尾素可能通过激活EBF3/ALOX15通路诱导A549细胞铁死亡,抑制细胞增殖与迁移。
Objective To investigate the effect of irisin regulating early B cytokine 3(EBF3)/arachidonic acid-15-lipoxygenase(ALOX15)pathway on the proliferation and migration of lung adenocarcinoma cells.Methods A549 cells were assigned into the irisin solvent group,the irisin group,the sh-NC group,the EBF3 inhibitor(sh-EBF3)group,the irisin+sh-NC group and the irisin+sh-EBF3 group randomly.5-bromo-2-deoxyuracil(EdU)staining and CCK-8 method were applied to detect cell proliferation.5-Scratch experiment was applied to detect the scratch healing rate.2′,7′-dichlorofluorescein diacetate(DCFH-DA)staining was applied to detect the level of reactive oxygen species(ROS)in cells.The reagent kit was used to detect glutathione(GSH),malondialdehyde(MDA)and ferrous ion(Fe^(2+))in cells.Transmission electron microscopy was applied to observe mitochondrial morphology in A549 cells.QRT-PCR was applied to detect mRNA levels of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase 2(MMP-2)and glutathione peroxidase 4(GPX4)in A549 cells.Western blot assay was applied to detect EBF3 and ALOX15 proteins in cells.Results Compared with the irisin solvent group,the mitochondria of A549 cells in the irisin group showed ferroptosis characteristics,the positive rate of EdU,OD_(450) value,scratch healing rate,GSH level,PCNA,MMP-2 and GPX4 mRNA levels decreased,and the ROS relative fluorescence intensity,MDA,Fe^(2+)level,and EBF3 and ALOX15 protein levels increased(P<0.05).Compared with the sh-NC group,the mitochondrial ferroptosis phenomenon of A549 cells was reduced in the sh-EBF3 group,the positive rate of EdU,OD_(450) value,scratch healing rate,GSH level,PCNA,MMP-2 and GPX4 mRNA levels increased,and the ROS relative fluorescence intensity,MDA,Fe^(2+)levels and EBF3 and ALOX15 protein levels reduced(P<0.05).Sh-EBF3 reversed the effect of irisin on ferroptosis,proliferation and migration of A549 cells.Conclusion Irisin may induce ferroptosis in A549 cells and inhibit cell proliferation and migration by activating the EBF3/ALOX15 pathway.
作者
苏红见
张春艳
张卫东
韩利
乔亚红
SU Hongjian;ZHANG Chunyan;ZHANG Weidong;HAN Li;QIAO Yahong(The Second Ward of Respiratory and Critical Care Department,Henan Provincial Chest Hospital,Zhengzhou 450000,China;the Third Ward of Respiratory and Critical Care Department,Henan Provincial Chest Hospital,Zhengzhou 450000,China;the Seventh Ward of Thoracic Surgery,Henan Provincial Chest Hospital,Zhengzhou 450000,China)
出处
《天津医药》
2025年第4期337-342,共6页
Tianjin Medical Journal
基金
河南省科技攻关项目(242102311079)。
