癌变·畸变·突变2017，Vol.29Issue(6)：411-417,7.DOI:10.3969/j.issn.1004-616x.2017.06.002
TGF-β1诱导上皮间质转化过程中线粒体合成与功能的改变
Alteration of mitochondrial synthesis and function in epithelial-mesenchymal transition by TGF-β1
摘要
Abstract
OBJECTIVE:To investigate changes of ROS and mitochondria on epithelial-mesenchymal transition.METHODS:Human lung cancer A549 cells were divided into control and treated groups.The latter were treated with different concentrations of TGF-β1 (0,2.5,5,10,20,40 ng/mL) and morphological changes were observed after 48 hours.In addition,cell viability was detected by using the CCK-8 kit and cell apoptosis by flow cytometry.DCFH fluorescent probe,MitoSOX fluorescent probe and Rhodamine-123 staining were used to detect ROS and changes in mitochondrial membrane potential,respectively.The content of ATP was detect by using the enhanced ATP kit.The epithelial-mesenchymal transition-related markers and mitochondrial-related proteins were detected by Western blot.RESULTS:The treated groups showed major differences from the control group:cell viability increased gradually and showed a dose-effect relationship with increased concentrations of TGF-β1 (r=-0.941,P＜0.05);there was a significant difference on cell apoptosis after treatment with 5-20 ng/mL TGF-β1;mesenchymal cells were observed after treatment with TGF-β1 for 48 hours,indicating the appearance of epithelial mesenchymal transition;Western blot showed that the expression of epithelial marker E-cadherin decreased gradually and the expression of mesenchymal marker α-SMA increased gradually (P＜0.05);the detection of intracellular reactive oxygen showed that treatment with 5-20 ng/mL TGF-β1 induced generation of ROS and mitochondrial ROS (P＜0.05);mitochondrial membrane potential declined as indicated by fluorescence intensity (P＜0.05);the content of ATP also decreased (P＜0.05);and expression of mitochondrial-related proteins were downregulated (P＜0.05).CONCLUSION:TGF-β1 caused increase of reactive oxygen species which led to the occurrence of mitochondrial dysfunction and promotion of EMT.关键词
上皮间质转化/活性氧/线粒体/转化生长因子-β1Key words
epithelial mesenchymal transition/reactive oxygen species/mitochondrial dysfunction分类
医药卫生引用本文复制引用
张佳欣,刘颖,张伟,周庆彪,孔德钦,刘瑞,海春旭..TGF-β1诱导上皮间质转化过程中线粒体合成与功能的改变[J].癌变·畸变·突变,2017,29(6):411-417,7.基金项目
国家自然科学基金(81573126) （81573126）
陕西省自然科学基金青年人才项目(2014JQ4141) （2014JQ4141）
