Abstract
Despite significant progress, our understanding of how specific oncogenes transform cells is still limited and likely underestimates the complexity of downstream signalling events. To address this gap, we use mass spectrometry-based chemical proteomics to characterize the global impact of an oncogene on the expressed kinome, and then functionally annotate the regulated kinases. As an example, we identify 63 protein kinases exhibiting altered expression and/or phosphorylation in Src-transformed mammary epithelial cells. An integrated siRNA screen identifies nine kinases, including SGK1, as being essential for Src-induced transformation. Accordingly, we find that Src positively regulates SGK1 expression in triple negative breast cancer cells, which exhibit a prominent signalling network governed by Src family kinases. Furthermore, combined inhibition of Src and SGK1 reduces colony formation and xenograft growth more effectively than either treatment alone. Therefore, this approach not only provides mechanistic insights into oncogenic transformation but also aids the design of improved therapeutic strategies.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antineoplastic Agents / pharmacology
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Antineoplastic Agents / therapeutic use
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Benzodioxoles / pharmacology
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Benzodioxoles / therapeutic use
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Cell Line, Tumor
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Cell Transformation, Neoplastic / drug effects
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Cell Transformation, Neoplastic / genetics*
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Cell Transformation, Neoplastic / pathology
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Female
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Gene Expression Regulation, Neoplastic*
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Gene Knockdown Techniques
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HEK293 Cells
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Humans
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Immediate-Early Proteins / antagonists & inhibitors
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Immediate-Early Proteins / metabolism*
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Mass Spectrometry / methods
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Mice, Inbred BALB C
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Mice, Nude
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Oncogenes / genetics
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Protein Interaction Mapping / methods
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Protein Serine-Threonine Kinases / metabolism*
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Proteomics / methods
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Quinazolines / pharmacology
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Quinazolines / therapeutic use
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RNA, Small Interfering / metabolism
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Signal Transduction / drug effects
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Signal Transduction / genetics
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Triple Negative Breast Neoplasms / drug therapy
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Triple Negative Breast Neoplasms / genetics*
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Triple Negative Breast Neoplasms / pathology
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Xenograft Model Antitumor Assays
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src-Family Kinases / antagonists & inhibitors
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src-Family Kinases / metabolism*
Substances
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Antineoplastic Agents
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Benzodioxoles
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Immediate-Early Proteins
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Quinazolines
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RNA, Small Interfering
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saracatinib
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src-Family Kinases
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Protein Serine-Threonine Kinases
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serum-glucocorticoid regulated kinase