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MCF7 response to EGF: Difference between revisions

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|series=IN_VITRO DIFFERENTIATION SERIES
|series=IN_VITRO DIFFERENTIATION SERIES
|species=Human (Homo sapiens)
|species=Human (Homo sapiens)
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|time_points=0hr
|time_points=0hr
|time_span=8 hours
|time_span=8 hours
|timepoint_design=Early focus
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|tissue_cell_type=Breast epithelia cells
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Latest revision as of 17:17, 14 March 2022

Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:Zenbu
Expression table:FILE
Link to TET:TET
Sample providers :Mariko Okada
Germ layer:mesoderm
Primary cells or cell line:cell line
Time span:8 hours
Number of time points:16


Overview

ErbB receptor family plays a central role in cellular development, proliferation, differentiation and cell death. There are four members, ErbB1/EGFR, ErbB2, ErbB3 and ErbB4 receptors, belong to this family. Dysregulation of the receptors is highly associated with incidence of variety of cancers [1]. Once activated by growth factor binding, the receptor forms homo- and heterodimer and are trans-activated by tyrosine phosphorylation, transmits the phosphorylation signal to downstream kinase such as ERK and Akt, then those kinases activate/phosphorylate transcription factors such as ELK1, CREB, or SRF in nucleus to initiate gene expression.In human breast cancer MCF-7 cells, the heregulin (HRG; a ErbB3/4 ligand) induces cell differentiation (accumulation of lipid droplets), while epidermal growth factor (EGF; an ErbB1/EGFR ligand) elicits cell proliferation after 5-14 days of the ligand stimulation.These ligand-stimulated cells show similar time-course profiles in terms of signaling activity and immediate early gene (IEG) mRNA expression (up to 1.5 hours)[2,3]. However, analysis of delayed transcription (up to 72 hours) using qRT-PCR showed the significant expression of transcription factors (c-FOS, FRA-1 and FHL2) specific to HRG but not by EGF [4], suggesting the existence of unknown transcription machinery for cell differentiation during this time-course.
Figure 1: Approximate expression timing of early, mid, delayed response genes.

References:
[1] Yarden Y1, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol. 2, 127-137, 2001.
[2] Nagashima, et al. Quantitative transcriptional control of ErbB receptor signaling undergoes graded to biphasic response for cell differentiation. J. Biol. Chem. 282, 4045-4056, 2007.
[3] Nakakuki, et al. Ligand-specific c-Fos expression emerges from the spatiotemporal control of ErbB network dynamics. Cell 141, 884-896, 2010.
[4] Saeki, et al. Ligand-specific sequential regulation of transcription factors for differentiation of MCF-7 cells. BMC Genomics 10, 545, 2009.

Sample description

The MCF-7 human breast cancer cell line was obtained from American Type Culture Collection (ATCC) and maintained in DMEM (Gibco BRL, Githersburg, MD) supplemented with 10 % fetal bovine serum. Prior to growth hormone treatment, the cells were serum-starved for 16-24 hours, and then EGF (PeproTech House, London, England) or HRG-176-246 (R&D Systems, Inc., Minneapolis, MN) was added. We prepared the EGF or HRG-stimulated time-course samples at 0 (non-treated), 15min, 30min, 45min, 60min, 80min, 100min, 2hr, 2.5hr, 3hr, 3.5hr, 4hr, 5hr, 6hr, 7hr and 8hr, to cover the early phase of cell differentiation and to cover early, mid and delayed gene expression. The all cell samples were snap frozen in liquid nitrogen and provided for RNA extraction for CAGE analysis.
MCF-7 cells were incubated in the absence or presence of growth factor in serum-free DMEM. Following cultivation for 14 days, cells were stained with Oil Red O to evaluate differentiation of the cells, which results in lipid droplets accumulation (The figures from Nagashima et al. JBC 2007 [2]).

References:
[2] Nagashima, et al. Quantitative transcriptional control of ErbB receptor signaling undergoes graded to biphasic response for cell differentiation. J. Biol. Chem. 282, 4045-4056, 2007.

Quality control

The mRNA expression of c-FOS, FOSL1 and FHL2 was significantly induced for HRG than EGF shown in the past study (The figures from Saeki et al BMC Genomics 2009[4]).

Time-course patterns the CAGE peaks of c-FOS, FOSL1, FHL2 obtained from current analysis was consistent with the mRNA expression data shown in above.

References:
[4] Saeki, et al. Ligand-specific sequential regulation of transcription factors for differentiation of MCF-7 cells. BMC Genomics 10, 545, 2009.

Profiled time course samples

Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples


13032-139E8MCF7 breast cancer cell line response to EGF100hr15minbiol_rep1
13033-139E9MCF7 breast cancer cell line response to EGF100hr30minbiol_rep1
13034-139F1MCF7 breast cancer cell line response to EGF100hr45minbiol_rep1
13035-139F2MCF7 breast cancer cell line response to EGF101hr00minbiol_rep1
13036-139F3MCF7 breast cancer cell line response to EGF101hr20minbiol_rep1
13037-139F4MCF7 breast cancer cell line response to EGF101hr40minbiol_rep1
13038-139F5MCF7 breast cancer cell line response to EGF102hr00minbiol_rep1
13039-139F6MCF7 breast cancer cell line response to EGF102hr30minbiol_rep1
13040-139F7MCF7 breast cancer cell line response to EGF103hr00minbiol_rep1
13041-139F8MCF7 breast cancer cell line response to EGF103hr30minbiol_rep1
13042-139F9MCF7 breast cancer cell line response to EGF104hrbiol_rep1
13043-139G1MCF7 breast cancer cell line response to EGF105hrbiol_rep1
13044-139G2MCF7 breast cancer cell line response to EGF106hrbiol_rep1
13045-139G3MCF7 breast cancer cell line response to EGF107hrbiol_rep1
13046-139G4MCF7 breast cancer cell line response to EGF108hrbiol_rep1
13097-140D1MCF7 breast cancer cell line response to EGF100hr00minbiol_rep2
13098-140D2MCF7 breast cancer cell line response to EGF100hr15minbiol_rep2
13099-140D3MCF7 breast cancer cell line response to EGF100hr30minbiol_rep2
13100-140D4MCF7 breast cancer cell line response to EGF100hr45minbiol_rep2
13101-140D5MCF7 breast cancer cell line response to EGF101hr00minbiol_rep2
13102-140D6MCF7 breast cancer cell line response to EGF101hr20minbiol_rep2
13103-140D7MCF7 breast cancer cell line response to EGF101hr40minbiol_rep2
13104-140D8MCF7 breast cancer cell line response to EGF102hr00minbiol_rep2
13105-140D9MCF7 breast cancer cell line response to EGF102hr30minbiol_rep2
13106-140E1MCF7 breast cancer cell line response to EGF103hr00minbiol_rep2
13107-140E2MCF7 breast cancer cell line response to EGF103hr30minbiol_rep2
13108-140E3MCF7 breast cancer cell line response to EGF104hrbiol_rep2
13109-140E4MCF7 breast cancer cell line response to EGF105hrbiol_rep2
13110-140E5MCF7 breast cancer cell line response to EGF106hrbiol_rep2
13111-140E6MCF7 breast cancer cell line response to EGF107hrbiol_rep2
13112-140E7MCF7 breast cancer cell line response to EGF108hrbiol_rep2
13163-141B4MCF7 breast cancer cell line response to EGF100hr00minbiol_rep3
13164-141B5MCF7 breast cancer cell line response to EGF100hr15minbiol_rep3
13165-141B6MCF7 breast cancer cell line response to EGF100hr30minbiol_rep3
13166-141B7MCF7 breast cancer cell line response to EGF100hr45minbiol_rep3
13167-141B8MCF7 breast cancer cell line response to EGF101hr00minbiol_rep3
13168-141B9MCF7 breast cancer cell line response to EGF101hr20minbiol_rep3
13169-141C1MCF7 breast cancer cell line response to EGF101hr40minbiol_rep3
13170-141C2MCF7 breast cancer cell line response to EGF102hr00minbiol_rep3
13171-141C3MCF7 breast cancer cell line response to EGF102hr30minbiol_rep3
13172-141C4MCF7 breast cancer cell line response to EGF103hr00minbiol_rep3
13173-141C5MCF7 breast cancer cell line response to EGF103hr30minbiol_rep3
13174-141C6MCF7 breast cancer cell line response to EGF104hrbiol_rep3
13175-141C7MCF7 breast cancer cell line response to EGF105hrbiol_rep3
13176-141C8MCF7 breast cancer cell line response to EGF106hrbiol_rep3
13177-141C9MCF7 breast cancer cell line response to EGF107hrbiol_rep3
13178-141D1MCF7 breast cancer cell line response to EGF108hrbiol_rep3
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