Keshamouni Venkateshwar G, Michailidis George, Grasso Catherine S, Anthwal Shalini, Strahler John R, Walker Angela, Arenberg Douglas A, Reddy Raju C, Akulapalli Sudhakar, Thannickal Victor J, Standiford Theodore J, Andrews Philip C, Omenn Gilbert S
Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Michigan Proteomics Consortium, National Resource for Proteomics and Pathways, University of Michigan, Ann Arbor, Michigan, 48109, USA.
J Proteome Res. 2006 May;5(5):1143-54. doi: 10.1021/pr050455t.
Transforming growth factor-beta (TGF-beta) induces epithelial-mesenchymal transition (EMT) of epithelial cells in both normal embryonic development and certain pathological contexts. Here, we show that TGF-beta induced-EMT in human lung cancer cells (A549; adenocarcinoma cells) mediates tumor cell migration and invasion phenotypes. To gain insights into molecular events during EMT, we employed a global stable isotope labeled profiling strategy using iTRAQ reagents, followed by 2DLC-MS/MS, which identified a total of 51 differentially expressed proteins during EMT; 29 proteins were up-regulated and 22 proteins were down-regulated. Down-regulated proteins were predominantly enzymes involved in regulating nutrient or drug metabolism. The majority of the TGF-beta-induced proteins (such as tropomyosins, filamin A, B, & C, integrin-beta1, heat shock protein27, transglutaminase2, cofilin, 14-3-3 zeta, ezrin-radixin-moesin) are involved in the regulation of cell migration, adhesion and invasion, suggesting the acquisition of a invasive phenotype.
在正常胚胎发育和某些病理情况下，转化生长因子-β（TGF-β）均可诱导上皮细胞发生上皮-间质转化（EMT）。在此，我们发现TGF-β诱导人肺癌细胞（A549；腺癌细胞）发生EMT，介导肿瘤细胞的迁移和侵袭表型。为深入了解EMT过程中的分子事件，我们采用了一种基于iTRAQ试剂的全稳定同位素标记分析策略，随后进行二维液相色谱-串联质谱分析（2DLC-MS/MS），共鉴定出EMT过程中51种差异表达蛋白；其中29种蛋白上调，22种蛋白下调。下调的蛋白主要是参与调节营养物质或药物代谢的酶。大多数TGF-β诱导的蛋白（如原肌球蛋白、细丝蛋白A、B和C、整合素-β1、热休克蛋白27、转谷氨酰胺酶2、丝切蛋白、14-3-3ζ、埃兹蛋白-根蛋白-膜突蛋白）参与细胞迁移、黏附和侵袭的调节，提示获得了侵袭性表型。
