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MCF7 response to HRG

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Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:Zenbu
Expression table:TET
Link to TET:FILE
Sample providers :Mariko Okada
Germ layer:ectoderm
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



13048-139G6MCF7 breast cancer cell line response to HRG00hr15minbiol_rep1
13049-139G7MCF7 breast cancer cell line response to HRG00hr30minbiol_rep1
13050-139G8MCF7 breast cancer cell line response to HRG00hr45minbiol_rep1
13051-139G9MCF7 breast cancer cell line response to HRG01hr00minbiol_rep1
13052-139H1MCF7 breast cancer cell line response to HRG01hr20minbiol_rep1
13053-139H2MCF7 breast cancer cell line response to HRG01hr40minbiol_rep1
13054-139H3MCF7 breast cancer cell line response to HRG02hr00minbiol_rep1
13055-139H4MCF7 breast cancer cell line response to HRG02hr30minbiol_rep1
13056-139H5MCF7 breast cancer cell line response to HRG03hr00minbiol_rep1
13057-139H6MCF7 breast cancer cell line response to HRG03hr30minbiol_rep1
13058-139H7MCF7 breast cancer cell line response to HRG04hrbiol_rep1
13059-139H8MCF7 breast cancer cell line response to HRG05hrbiol_rep1
13060-139H9MCF7 breast cancer cell line response to HRG06hrbiol_rep1
13061-139I1MCF7 breast cancer cell line response to HRG07hrbiol_rep1
13062-139I2MCF7 breast cancer cell line response to HRG08hrbiol_rep1
13097-140D1MCF7 breast cancer cell line response to EGF100hr00minbiol_rep2
13114-140E9MCF7 breast cancer cell line response to HRG00hr15minbiol_rep2
13115-140F1MCF7 breast cancer cell line response to HRG00hr30minbiol_rep2
13116-140F2MCF7 breast cancer cell line response to HRG00hr45minbiol_rep2
13117-140F3MCF7 breast cancer cell line response to HRG01hr00minbiol_rep2
13118-140F4MCF7 breast cancer cell line response to HRG01hr20minbiol_rep2
13119-140F5MCF7 breast cancer cell line response to HRG01hr40minbiol_rep2
13120-140F6MCF7 breast cancer cell line response to HRG02hr00minbiol_rep2
13121-140F7MCF7 breast cancer cell line response to HRG02hr30minbiol_rep2
13122-140F8MCF7 breast cancer cell line response to HRG03hr00minbiol_rep2
13123-140F9MCF7 breast cancer cell line response to HRG03hr30minbiol_rep2
13124-140G1MCF7 breast cancer cell line response to HRG04hrbiol_rep2
13125-140G2MCF7 breast cancer cell line response to HRG05hrbiol_rep2
13126-140G3MCF7 breast cancer cell line response to HRG06hrbiol_rep2
13127-140G4MCF7 breast cancer cell line response to HRG07hrbiol_rep2
13128-140G5MCF7 breast cancer cell line response to HRG08hrbiol_rep2
13163-141B4MCF7 breast cancer cell line response to EGF100hr00minbiol_rep3
13180-141D3MCF7 breast cancer cell line response to HRG00hr15minbiol_rep3
13181-141D4MCF7 breast cancer cell line response to HRG00hr30minbiol_rep3
13182-141D5MCF7 breast cancer cell line response to HRG00hr45minbiol_rep3
13183-141D6MCF7 breast cancer cell line response to HRG01hr00minbiol_rep3
13184-141D7MCF7 breast cancer cell line response to HRG01hr20minbiol_rep3
13185-141D8MCF7 breast cancer cell line response to HRG01hr40minbiol_rep3
13186-141D9MCF7 breast cancer cell line response to HRG02hr00minbiol_rep3
13187-141E1MCF7 breast cancer cell line response to HRG02hr30minbiol_rep3
13188-141E2MCF7 breast cancer cell line response to HRG03hr00minbiol_rep3
13189-141E3MCF7 breast cancer cell line response to HRG03hr30minbiol_rep3
13190-141E4MCF7 breast cancer cell line response to HRG04hrbiol_rep3
13191-141E5MCF7 breast cancer cell line response to HRG05hrbiol_rep3
13192-141E6MCF7 breast cancer cell line response to HRG06hrbiol_rep3
13193-141E7MCF7 breast cancer cell line response to HRG07hrbiol_rep3
13194-141E8MCF7 breast cancer cell line response to HRG08hrbiol_rep3