Abstract
In this study, we have addressed how Lyn kinase signaling mediates nilotinib-resistance by quantitative phospho-proteomics using Stable Isotope Labeling with Amino acid in Cell culture. We have found an increased tyrosine phosphorylation of 2 additional tyrosine kinases in nilotinib-resistant cells: the spleen tyrosine kinase Syk and the UFO family receptor tyrosine kinase Axl. This increased tyrosine phosphorylation involved an interaction of these tyrosine kinases with Lyn. Inhibition of Syk by the inhibitors R406 or BAY 61-3606 or by RNA interference restored the capacity of nilotinib to inhibit cell proliferation. Conversely, coexpression of Lyn and Syk were required to fully induce resistance to nilotinib in drug-sensitive cells. Surprisingly, the knockdown of Syk also strongly decreased tyrosine phosphorylation of Lyn and Axl, thus uncovering interplay between Syk and Lyn. We have shown the involvement of the adaptor protein CDCP-1 in resistance to nilotinib. Interestingly, the expression of Axl and CDCP1 were found increased both in a nilotinib-resistant cell line and in nilotinib-resistant CML patients. We conclude that an oncogenic signaling mediated by Lyn and Syk can bypass the need of Bcr-Abl in CML cells. Thus, targeting these kinases may be of therapeutic value to override imatinib or nilotinib resistance in CML.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, CD / genetics
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Antigens, CD / metabolism
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Antigens, Neoplasm
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Antineoplastic Agents / pharmacology*
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Axl Receptor Tyrosine Kinase
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Base Sequence
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Cell Adhesion Molecules / genetics
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Cell Adhesion Molecules / metabolism
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Drug Resistance, Neoplasm / genetics
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Drug Resistance, Neoplasm / physiology
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Female
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Fusion Proteins, bcr-abl / metabolism
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Gene Expression
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Humans
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Intracellular Signaling Peptides and Proteins / antagonists & inhibitors
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Intracellular Signaling Peptides and Proteins / genetics
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Intracellular Signaling Peptides and Proteins / metabolism*
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K562 Cells
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive / drug therapy*
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive / metabolism*
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Male
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Neoplasm Proteins / genetics
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Neoplasm Proteins / metabolism
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Phosphorylation
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Protein-Tyrosine Kinases / antagonists & inhibitors
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Protein-Tyrosine Kinases / genetics
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Protein-Tyrosine Kinases / metabolism*
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Proteomics
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Proto-Oncogene Proteins / metabolism
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Pyrimidines / pharmacology*
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RNA, Small Interfering / genetics
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Receptor Protein-Tyrosine Kinases / metabolism
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Signal Transduction
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Syk Kinase
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src-Family Kinases / genetics
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src-Family Kinases / metabolism*
Substances
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Antigens, CD
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Antigens, Neoplasm
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Antineoplastic Agents
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CDCP1 protein, human
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Cell Adhesion Molecules
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Intracellular Signaling Peptides and Proteins
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Neoplasm Proteins
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Proto-Oncogene Proteins
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Pyrimidines
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RNA, Small Interfering
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Protein-Tyrosine Kinases
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Receptor Protein-Tyrosine Kinases
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Fusion Proteins, bcr-abl
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SYK protein, human
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Syk Kinase
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lyn protein-tyrosine kinase
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src-Family Kinases
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nilotinib
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Axl Receptor Tyrosine Kinase
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AXL protein, human