Temporal Quantitative Proteomics by iTRAQ 2D-LC-MS/MS and Corresponding mRNA Expression Analysis Identify Post-Transcriptional Modulation of Actin-Cytoskeleton Regulators During TGF-β-Induced Epi
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- 作者：Venkateshwar G. Keshamouni ; Pratik Jagtap ; George Michailidis ; John R. Strahler ; Rork Kuick ; Ajaya Kumar Reka ; Panagiotis Papoulias ; Rashmi Krishnapuram ; Anjaiah Srirangam ; Theodore J. Standiford ; Phi
- 刊名：Journal of Proteome Research
- 出版年：2009
- 出版时间：January 2, 2009
- 年：2009
- 卷：8
- 期：1
- 页码：35-47
- 全文大小：331K
- 年卷期：v.8,no.1(January 2, 2009)
- ISSN：1535-3907
文摘
To gain insights into how TGF-β regulates epithelial-mesenchymal transition (EMT), we assessed the time course of proteins and mRNAs during EMT by multiplex iTRAQ labeling and 2D-LC-MS/MS, and by hybridization, respectively. Temporal iTRAQ analysis identified 66 proteins as differentially expressed during EMT, including newly associated proteins calpain, fascin and macrophage-migration inhibitory factor (MIF). Comparing protein and mRNA expression overtime showed that all the 14 up-regulated proteins involved in the actin-cytoskeleton remodeling were accompanied by increases in corresponding mRNA expression. Interestingly, siRNA mediated knockdown of cofilin1 potentiated TGF-β-induced EMT. Further analysis of cofilin1 and β-actin revealed an increase in their mRNA stability in response to TGF-β, contributing to the observed increase in mRNA and protein expression. These results are the first demonstration of post-transcriptional regulation of cytoskeletal remodelling and a key role for cofilin1 during TGF-β-induced EMT.