Phosphoproteomic Study of Human Tubular Epithelial Cell in Response to Transforming Growth Factor-Beta-1-Induced Epithelial-to-Mesenchymal Transition

被引：24

作者：

Chen, Yong-Xi ^{[1
]}

Li, Ya ^{[1
]}

Wang, Wei-Ming ^{[1
]}

Zhang, Wen ^{[1
]}

Chen, Xiao-Nong ^{[1
]}

Xie, Yin-Yin ^{[2
,3
]}

Lu, Jing ^{[2
,3
]}

Huang, Qiu-Hua ^{[2
,3
]}

Chen, Nan ^{[1
]}

机构：

[1] Shanghai Jiao Tong Univ, Dept Nephrol, Shanghai Ruijin Hosp, Sch Med, Shanghai 200025, Peoples R China

[2] Shanghai Jiao Tong Univ, State Key Lab Med Genom, Shanghai Ruijin Hosp, Sch Med, Shanghai 200025, Peoples R China

[3] Shanghai Jiao Tong Univ, Shanghai Inst Hematol, Shanghai Ruijin Hosp, Sch Med, Shanghai 200025, Peoples R China

来源：

AMERICAN JOURNAL OF NEPHROLOGY | 2010年 / 31卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Isobaric tags for relative and absolute quantification; Transforming growth factor-beta; Tubular epithelial cell; Epithelial-to-mesenchymal transition; Chronic kidney disease; 2-DIMENSIONAL GEL-ELECTROPHORESIS; ACTIVATED PROTEIN-KINASE; TGF-BETA; PROTEOMIC ANALYSIS; ERM PROTEINS; ACTIN-CYTOSKELETON; URINARY PROTEOME; PHOSPHORYLATION; PATHOGENESIS; EXPRESSION;

D O I：

10.1159/000253865

中图分类号：

R5 [内科学]; R69 [泌尿科学（泌尿生殖系疾病）];

学科分类号：

1002 ; 100201 ;

摘要：

Background: Transforming growth factor-beta (TGF-beta)-induced epithelial-to-mesenchymal transition (EMT) plays an important role in renal fibrosis and progression of chronic kidney disease (CKD). Phosphorylation of proteins is essential to TGF-beta signaling. We applied isobaric tags for relative and absolute quantification (iTRAQ) technology to profile the phosphoproteins in tubular epithelial cells in response to TGF-beta-induced EMT in order to further study molecular events. Methods: HK-2 cells were treated with TGF-beta 1 to induce EMT. The cells were divided into a control group (without TGF-beta 1 treatment) and a TGF-beta 1-treated group. Phosphoproteins from two groups were extracted and differentially labeled with iTRAQ reagents and processed by 2D-nano-HPLC-MS/MS. Validating of iTRAQ analysis was performed by western blot. Bioinformatic analysis was performed by on-line databases. Results: By iTRAQ-2D-nano-HPLC-MS/MS, 38 differentially expressed phosphoproteins were identified which included 19 up-regulated phosphoproteins and 19 down-regulated phosphoproteins. Western blot confirmed up-regulation of phosphorylated moesin and HSP90 alpha. Bioinformatic analysis suggested that the majority of proteins were located in the nucleus and endoplasmic reticulum lumen. The phosphoproteins were categorized into 17 molecular function classifications. Nucleic acid binding protein, cytoskeletal protein and chaperone were the major categories of molecular function. A biological network was built to analyze interaction between up-regulated proteins. Conclusion: We demonstrate a TGF-beta 1-mediated post-transcriptional regulation of EMT in tubular epithelial cells. Phosphorylation of moesin and HSP90 alpha might play a role in TGF-beta-induced EMT. Copyright (c) 2009 S. Karger AG, Basel

引用

页码：24 / 35

页数：12

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