Dataset Information

Phosphoproteome of MCF7 breast cancer cells treated with estradiol and BI2536

ABSTRACT: Triple SILAC phosphoproteome experiment of hormone deprived MCF7 cells treated with estradiol (E2) in presence or absence of the PLK1 inhibitor BI2536. Conditions were as follows: light (L) population: 1h DMSO, 30min EtOH; medium (M) population: 1h DMSO, 30min E2; heavy (H) population: 1h BI2536, 30min E2. Biological replicates: A and B. Phosphopeptides were enriched from soluble and insoluble fraction using a two step enrichment protocol consiting of strong cation exchange and TiO2 affinity chromatography. The peptides were analyzed on an Orbitrap Velos mass spectrometer linked to an online nanoflow HPLC system via a nanoelectrospray ion source and fragmented via higher-energy C-trap dissociation (HCD). Analysis was carried out using MaxQuant (v. 1.2.2.5) with standard settings including STY phosphporylation as variable modification.

OTHER RELATED OMICS DATASETS IN: PRJNA202861

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Breast Cancer Cell

SUBMITTER: Michael Wierer

LAB HEAD: Miguel Beato

PROVIDER: PXD000275 | Pride | 2016-06-24

REPOSITORIES: Pride

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Publications

PLK1 signaling in breast cancer cells cooperates with estrogen receptor-dependent gene transcription.

Wierer Michael M Verde Gaetano G Pisano Paola P Molina Henrik H Font-Mateu Jofre J Di Croce Luciano L Beato Miguel M

Cell reports 20130613 6

Polo-like kinase 1 (PLK1) is a key regulator of cell division and is overexpressed in many types of human cancers. Compared to its well-characterized role in mitosis, little is known about PLK1 functions in interphase. Here, we report that PLK1 mediates estrogen receptor (ER)-regulated gene transcription in human breast cancer cells. PLK1 interacts with ER and is recruited to ER cis-elements on chromatin. PLK1-coactivated genes included classical ER target genes such as Ps2, Wisp2, and Serpina3 ...[more]

PMID: 23770244

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Project description:Developmental exposure of mouse fetuses to estrogens results in dose-dependent permanent effects on prostate morphology and function. Fetal prostatic mesenchyme cells express estrogen receptor alpha (ERα) and androgen receptors and convert stimuli from estrogens and androgens into signaling to regulate epithelial cell proliferation and differentiation. To obtain mechanistic insight into the role of different doses of estradiol (E2) in regulating mesenchymal cells, we examined E2-induced transcriptomal changes in primary cultures of fetal mouse prostate mesenchymal cells. Urogenital sinus mesenchyme cells were obtained from male mouse fetuses at gestation day 17 and exposed to 10 pM, 100 pM or 100 nM E2 in the presence of a physiological concentration of dihydrotestosterone (0.69 nM) for four days. Gene ontology studies suggested that low doses of E2 (10 pM and 100 pM) induce genes involved in cell adhesion, morphological tissue development, and sterol biosynthesis but suppress genes involved in growth factor signaling and cell adhesion. Genes showing inverted-U-shape dose responses (enhanced by E2 at 10 pM E2 but suppressed at 100 pM) were identified, and their enrichment in the glycolytic pathway was demonstrated. At the highest dose (100 nM), E2 induced genes enriched not only for cell adhesion but also steroid hormone signaling and metabolism, cytokines and their receptors, cell-to-cell communication, Wnt signaling, and TGF-β signaling. These results suggest that prostate mesenchymal cells may regulate epithelial cells through direct cell contacts when estrogen level is low whereas soluble growth factors might play significant roles when estrogen level is high. Primary culture urogenital sinus mesenchymal cells were isolated from prostate glands of gestation day 17 CD1 male mouse fetuses. Cells were then exposed to 10 pM or 100 pM 17beta-estradiol or vehicle (0.05% ethanol) for four days in the presence of 690 pM 5alpha-dihydrotestosterone. Total cellular RNA was then isolated for determination of transcriptomal profiles by Affymetrix mouse gene 1.0 ST array.

Dataset Information

Phosphoproteome of MCF7 breast cancer cells treated with estradiol and BI2536

Publications

PLK1 signaling in breast cancer cells cooperates with estrogen receptor-dependent gene transcription.

Similar Datasets

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