title: Quantitative Analysis of Differential Proteome Expression in Epithelial-to-Mesenchymal Transition of Bladder Epithelial Cells Using SILAC Method.
abstract: Epithelial-to-mesenchymal transition (EMT) is an essential biological process involved in embryonic development, cancer progression, and metastatic diseases. EMT has often been used as a model for elucidating the mechanisms that underlie bladder cancer progression. However, no study to date has addressed the quantitative global variation of proteins in EMT using normal and non-malignant bladder cells. We treated normal bladder epithelial HCV29 cells and low grade nonmuscle invasive bladder cancer KK47 cells with transforming growth factor-beta (TGF-β) to establish an EMT model, and studied non-treated and treated HCV29 and KK47 cells by the stable isotope labeling amino acids in cell culture (SILAC) method. Labeled proteins were analyzed by 2D ultrahigh-resolution liquid chromatography/LTQ Orbitrap mass spectrometry. Among a total of 2994 unique identified and annotated proteins in HCV29 and KK47 cells undergoing EMT, 48 and 56 proteins, respectively, were significantly upregulated, and 106 and 24 proteins were significantly downregulated. Gene ontology (GO) term analysis and pathways analysis indicated that the differentially regulated proteins were involved mainly in enhancement of DNA maintenance and inhibition of cell-cell adhesion. Proteomes were compared for bladder cell EMT vs. bladder cancer cells, revealing 16 proteins that displayed similar changes in the two situations. Studies are in progress to further characterize these 16 proteins and their biological functions in EMT.
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry &amp; Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. glyanglife@jiangnan.edu.cn.
authors.forename: Ganglong
authors.initials: G
authors.lastname: Yang
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry &amp; Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. luyuweifeng@gmail.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Lu
authors.affiliation: Wuxi Medical School, Jiangnan University, Wuxi 214122, China. jndxyd1988@163.com.
authors.forename: Di
authors.initials: D
authors.lastname: Yu
authors.affiliation: Department of Urology, Affiliated Hospital of Jiangnan University, Wuxi 214062, China. chengwensun@163.com.
authors.forename: Chengwen
authors.initials: C
authors.lastname: Sun
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry &amp; Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. guojia@jiangnan.edu.cn.
authors.forename: Jia
authors.initials: J
authors.lastname: Guo
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an 710069, China. zhengli@nwu.edu.cn.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry &amp; Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. fengguan@jiangnan.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Guan
mesh_terms: D002448:Cell Adhesion; D002460:Cell Line; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D004273:DNA, Neoplasm / Q000235:genetics* / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects* / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D063990:Gene Ontology; D053263:Gene Regulatory Networks / Q000187:drug effects; D006801:Humans; D007553:Isotope Labeling; D013058:Mass Spectrometry; D058977:Molecular Sequence Annotation; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism / Q000473:pathology
chemical_list: D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D020543:Proteome; D016212:Transforming Growth Factor beta
keywords: SILAC; bladder cancer; epithelial-to-mesenchymal transition (EMT); mass spectrometry; quantitative proteomics