TGF-β1诱导上皮间质转化过程中线粒体合成与功能的改变

doi:10.3969/j.issn.1004-616x.2017.06.002

Abstract

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.

Key words: epithelial mesenchymal transition, reactive oxygen species, mitochondrial dysfunction

CLC Number:

R73-37

ZHANG Jiaxin, LIU Ying, ZHANG Wei, ZHOU Qingbiao, KONG Deqin, LIU Rui, HAI Chunxu. Alteration of mitochondrial synthesis and function in epithelial-mesenchymal transition by TGF-β1[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(6): 411-417.

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Alteration of mitochondrial synthesis and function in epithelial-mesenchymal transition by TGF-β1

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