title: Impact of the mitogen-activated protein kinase pathway on the subproteome of detergent-resistant microdomains of colon carcinoma cells.
abstract: Lipid rafts play a key role in the regulation of fundamentally important cellular processes, including cell proliferation, differentiation, and survival. The composition of such detergent-resistant microdomains (DRMs) is altered under pathologic conditions, including cancer. Although DRMs have been analyzed in colorectal carcinoma little information exists about their composition upon treatment with targeted drugs. Hence, a quantitative proteomic profiling approach was performed to define alterations within the DRM fraction of colorectal carcinoma cells upon treatment with the drug U0126, an inhibitor of the mitogen-activated protein kinase pathway. Comparative expression profilings resulted in the identification of 300 proteins, which could partially be linked to key oncogenic signaling pathways and tumor-related cellular features, such as cell proliferation, adhesion, motility, invasion, and apoptosis resistance. Most of these proteins were downregulated upon inhibitor treatment. In addition, quantitative proteomic profilings of cholesterol-depleted versus intact lipid rafts were performed to define, which U0126-regulated target structures represent bona fide raft proteins. Selected differentially abundant raft proteins were validated at the mRNA and/or protein level using U0126- or Trametinib-treated cells. The presented data provide insights into the molecular mechanisms associated with the response to the treatment with MEK inhibitors and might also lead to novel candidates for therapeutic interventions.
authors.affiliation: Institute of Medical Immunology, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
authors.forename: Christian V
authors.initials: CV
authors.lastname: Recktenwald
authors.forename: Rudolf
authors.initials: R
authors.lastname: Lichtenfels
authors.forename: Jens
authors.initials: J
authors.lastname: Wulfaenger
authors.forename: Anja
authors.initials: A
authors.lastname: Müller
authors.forename: Sven P
authors.initials: SP
authors.lastname: Dressler
authors.forename: Barbara
authors.initials: B
authors.lastname: Seliger
mesh_terms: D002070:Butadienes / Q000494:pharmacology; D045744:Cell Line, Tumor; D003106:Colon / Q000187:drug effects / Q000378:metabolism; D003110:Colonic Neoplasms / Q000378:metabolism*; D003902:Detergents / Q000378:metabolism; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002070:Butadienes; D003902:Detergents; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D020543:Proteome; C113580:U 0126; D020928:Mitogen-Activated Protein Kinases
keywords: Cell biology; Colon carcinoma; ICPL; LC-MALDI-MS; Lipid raft