title: Determination of differentially regulated proteins upon proteasome inhibition in AML cell lines by the combination of large-scale and targeted quantitative proteomics.
abstract: The ubiquitin-proteasome pathway (UPP) plays a critical role in the degradation of proteins implicated in cell cycle control, signal transduction, DNA damage response, apoptosis and immune response. Proteasome inhibitors can inhibit the growth of a broad spectrum of human cancer cells by altering the balance of intracellular proteins. However, the targets of these compounds in acute myeloid leukemia (AML) cells have not been fully characterized. Herein, we combined large-scale quantitative analysis by SILAC-MS and targeted quantitative proteomic analysis in order to identify proteins regulated upon proteasome inhibition in two AML cell lines displaying different stages of maturation: immature KG1a cells and mature U937 cells. In-depth data analysis enabled accurate quantification of more than 7000 proteins in these two cell lines. Several candidates were validated by selected reaction monitoring (SRM) measurements in a large number of samples. Despite the broad range of proteins known to be affected by proteasome inhibition, such as heat shock (HSP) and cell cycle proteins, our analysis identified new differentially regulated proteins, including IL-32, MORF family mortality factors and apoptosis inducing factor SIVA, a target of p53. It could explain why proteasome inhibitors induce stronger apoptotic responses in immature AML cells.
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Mariette
authors.initials: M
authors.lastname: Matondo
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Marlène
authors.initials: M
authors.lastname: Marcellin
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Karima
authors.initials: K
authors.lastname: Chaoui
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Bousquet-Dubouch
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Gonzalez-de-Peredo
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Monsarrat
authors.affiliation: Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Burlet-Schiltz
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D000069286:Bortezomib / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D019295:Computational Biology; D020869:Gene Expression Profiling; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects*; D063990:Gene Ontology; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007378:Interleukins / Q000235:genetics / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007976:Leupeptins / Q000494:pharmacology; D058977:Molecular Sequence Annotation; D010766:Phosphorylation / Q000187:drug effects; D046988:Proteasome Endopeptidase Complex / Q000187:drug effects* / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000494:pharmacology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D006360:Heat-Shock Proteins; C496971:IL32 protein, human; D007378:Interleukins; D007976:Leupeptins; C109882:MORF4 protein, human; D061988:Proteasome Inhibitors; D020033:Protein Isoforms; C106549:SIVA1 protein, human; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; C067713:lactacystin; D000069286:Bortezomib; D046988:Proteasome Endopeptidase Complex; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; D000111:Acetylcysteine
keywords: Apoptosis; Human acute myeloid leukemia (AML) cells; Proteasome inhibitor; SILAC; Targeted proteomics; Technology