Quantitative Analysis of Differential Proteome Expression in Epithelial-to-Mesenchymal Transition of Bladder Epithelial Cells Using SILAC Method

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Ganglong YangFeng Guan

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 ...Continue Reading

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Citations

Aug 21, 2019·Molecular Omics·Virgínia Campos SilvestriniVítor Marcel Faça
Dec 18, 2019·The Journal of Pharmacology and Experimental Therapeutics·Jun WangZhankui Jia
Apr 11, 2017·International Journal of Oncology·Jing-Bo ShaoZhi-Bao Lu
Dec 3, 2020·International Journal of Molecular Sciences·Shu-Ching OuBing-Chang Chen
Aug 8, 2021·Journal of Clinical Medicine·Margherita SistoSabrina Lisi

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Methods Mentioned

BETA
electrophoresis
glycosylation

Software Mentioned

Ingenuity Pathways Analysis ( IPA )
MaxQuant
Expression Analysis Systematic Explorer ( EASE )
SILAC
DAVID
Andromeda
Ingenuity Pathways Knowledge Base (

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