Characterization of the Tyrosine Kinase-Regulated Proteome in Breast Cancer by Combined use of RNA interference (RNAi) and Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) Quantitative Proteomics
File(s)Mol Cell Proteomics-2015-Stebbing-2479-92.pdf (5.16 MB)
Published version
Author(s)
Type
Journal Article
Abstract
Tyrosine kinases (TKs) are central regulators in cellular activities and perturbations of TK signaling contribute to oncogenesis. However, less than half of the TKs have been thoroughly studied and a global functional analysis of their proteomic portrait is lacking. Here we conducted a combined approach of RNA interference (RNAi) and stable isotope labeling with amino acids in cell culture (SILAC)-based quantitative proteomics to decode the TK-regulated proteome and associated signaling dynamics. As a result, a broad proteomic repertoire modulated by TKs was revealed, upon silencing of the 65 TKs expressed in MCF7 breast cancer cells. This yielded 10 new distinctive TK clusters according to similarity in TK-regulated proteome, each characterized by a unique signaling signature in contrast to previous classifications. We provide functional analyses and identify critical pathways for each cluster based on their common downstream targets. Analysis of different breast cancer subtypes showed distinct correlations of each cluster with clinical outcome. From the significantly up- and down-regulated proteins, we identified a number of markers of drug sensitivity and resistance. These data supports the role of TKs in regulating major aspects of cellular activity, but also reveals redundancy in signaling, explaining why kinase inhibitors alone often fail to achieve their clinical aims. The TK-SILACepedia provides a comprehensive resource for studying the global function of TKs in cancer.
Date Issued
2015-09-01
Date Acceptance
2015-06-13
Citation
Molecular & Cellular Proteomics, 2015, 14 (9), pp.2479-2492
ISSN
1535-9484
Publisher
American Society for Biochemistry and Molecular Biology
Start Page
2479
End Page
2492
Journal / Book Title
Molecular & Cellular Proteomics
Volume
14
Issue
9
Copyright Statement
© 2015 by The American Society for Biochemistry and Molecular Biology, Inc. Final version free via Creative Commons CC-BY license.
License URL
Sponsor
National Institute for Health Research
Cancer Research UK
Grant Number
NIHR-RP-011-053
C27532/A14549
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
GENE-EXPRESSION
DRUG-SENSITIVITY
DNA-REPLICATION
IN-VIVO
IDENTIFICATION
SCREEN
INVASION
NETWORKS
DYNAMICS
THERAPY
Publication Status
Published