A Proteomic Analysis Reveals That Snail Regulates the Expression of the Nuclear Orphan Receptor Nuclear Receptor Subfamily 2 Group F Member 6 (Nr2f6) and Interleukin 17 (IL-17) to Inhibit Adipocyte Differentiation *

Peláez‐García, Alberto; Barderas, Rodrigo; Batlle, Raquel; Viñas-Castells, Rosa; Bartolomé, Rubén A.; Torres, Sofía; Mendes, Marta; López-Lucendo, María; Mazzolini, Rocco; Bonilla, Félix; Herreros, Antonio Garcı́a de; Casal, J. Ignacio

doi:10.1074/mcp.m114.045328

Cited by 38 publications

(36 citation statements)

References 48 publications

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“…Despite several thousand publications linking Snail expression to cancer, at the time of this submission, there has been no direct mass spectrometry-based profile of the proteome-wide consequences of Snail over-expression in epithelial cells. A recent proteomic analysis of Snail-transduced adipocyte nuclear fractions identified 463 decreased and 111 increased proteins changed by more than 150% 28 . Extending from this analysis, human basal epithelial MCF10A cells were retrovirally transduced with Snail and labeled in isotopic SILAC media for quantitative proteomic analysis.…”

Section: Resultsmentioning

confidence: 99%

Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition

et al. 2016

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Epithelial cells form spatially-organized adhesion complexes that establish polarity gradients, regulate cell proliferation, and direct wound healing. As cells accumulate oncogenic mutations, these key tumor suppression mechanisms are disrupted, eliminating many adhesion complexes and bypassing contact inhibition. The transcription factor Snail is often expressed in malignant cancers, where it promotes transcriptional reprogramming to drive epithelial-mesenchymal transition (EMT), which promotes a more invasive state. S-palmitoylation describes the fatty-acyl post-translational modification of cysteine residues in proteins, and is required for membrane anchoring, trafficking, localization and function of hundreds of proteins involved in cell growth, polarity, and signaling. Since Snail-expression prevents apico-basolateral cell polarity, we asked if Snail-dependent transformation induces proteome-wide changes in S-palmitoylation. MCF10A breast cancer cells were retrovirally transduced with Snail, and correlated proteome-wide changes in protein abundance and S-palmitoylation were profiled using by stable isotope labeling in cell culture with amino acids (SILAC) mass spectrometry. This analysis identified increased levels of proteins involved in migration, glycolysis, and cell junction remodeling, and decreased levels of proteins involved in cell adhesion. Overall, protein S-palmitoylation is highly correlated with protein abundance, yet for a subset of proteins, this correlation is uncoupled. These findings suggest that Snail-overexpression affects the S-palmitoylation cycle of some proteins, and may affect cell polarity and tumor suppression.

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Section: Resultsmentioning

confidence: 99%

Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition

et al. 2016

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“…Total protein concentration of the resultant cellular lysates and tissue extracts were estimated using a BCA protein assay kit (Pierce, Rockford, IL) , whereas semi‐quantitative determinations were carried out using dot‐blot analysis . Details about procedures and assessment by blot‐assays of IL‐13Rα2 and E‐CDH status in cell lysates and paired N/T tissue extracts were given in previous works on the single determination of each biomarker .…”

Section: Methodsmentioning

confidence: 99%

Dual Amperometric Immunosensor for Improving Cancer Metastasis Detection by the Simultaneous Determination of Extracellular and Soluble Circulating Fraction of Emerging Metastatic Biomarkers

et al. 2019

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This paper reports the development of a dual immunosensor using magnetic microcarriers (MBs) and amperometric transduction at dual screen‐printed carbon electrodes (SPdCEs) for the simultaneous determination of two biomarkers: interleukin‐13 receptor α2 (IL‐13Rα2) and E‐cadherin (E‐CDH), with both extracellular and soluble fraction; oncogenic and tumor suppressor markers, respectively, of great relevance in metastatic processes. The implemented methodology involved the formation of sandwich‐type immunocomplexes using specific capture antibodies immobilized onto carboxylic acid magnetic microbeads (HOOC‐MBs), and biotinylated detector antibodies labeled with streptavidin−horseradish peroxidase conjugates (Strep‐HRP). The amperometric detection was performed by addition of hydrogen peroxide in the presence of hydroquinone (HQ) as the redox mediator. The dual immunosensing platform provided linear calibration ranges suitable for the determination of both biomarkers in liquid and solid clinical specimens as well as excellent selectivity against other cancer biomarkers. This simple handling dual bioplatform was applied to the determination of the soluble and extracellular fraction of the target biomarkers in serum and paraffined‐embedded tissues from colorectal cancer (CRC) patients diagnosed at different tumor grade. The obtained results reveal great potential of this configuration to improve the reliability in diagnosing metastatic CRC.

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“…Firstly, 40 μg of each extract replicate were separated by 15% SDS-PAGE and stained with Coomassie Brilliant Blue G 250 (Merck, Darmstadt, Germany). Then, whole lanes were cut into four (replicate 1) or six slices (replicates 2 and 3) prior to in-gel trypsin digestion (27,28).…”

Section: Sds-page Trypsin Digestion and Lc-ms/ms Analysismentioning

confidence: 99%

Delineation of the Olive Pollen Proteome and Its Allergenome Unmasks Cyclophilin as a Relevant Cross-Reactive Allergen

et al. 2019

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Olive pollen is a major allergenic source worldwide for its extensive cultivation. We have combined available genomic data with a comprehensive proteomic approach to get the annotated olive tree (Olea europaea L.) pollen proteome and define its complex allergenome. A total of 1,907 proteins were identified by LC-MS/MS using predicted protein sequences from its genome. Most proteins (60%) were predicted to possess catalytic activity and be involved in metabolic processes. In total, 203 proteins belonging to 47 allergen families were found in olive pollen. A peptidyl-prolyl cis-trans isomerase-cyclophilin-produced in Escherichia coli, was found as a new olive pollen allergen (Ole e 15). Most Ole e 15-sensitized patients were children (63%) and showed strong IgE recognition to the allergen. Ole e 15 shared high sequence identity with other plant, animal and fungal cyclophilins and presented high IgE cross-reactivity with pollen, plant food and animal extracts.

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A Proteomic Analysis Reveals That Snail Regulates the Expression of the Nuclear Orphan Receptor Nuclear Receptor Subfamily 2 Group F Member 6 (Nr2f6) and Interleukin 17 (IL-17) to Inhibit Adipocyte Differentiation *

Cited by 38 publications

References 48 publications

Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition

Correlated S-palmitoylation profiling of Snail-induced epithelial to mesenchymal transition

Dual Amperometric Immunosensor for Improving Cancer Metastasis Detection by the Simultaneous Determination of Extracellular and Soluble Circulating Fraction of Emerging Metastatic Biomarkers

Delineation of the Olive Pollen Proteome and Its Allergenome Unmasks Cyclophilin as a Relevant Cross-Reactive Allergen

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