MCF7 response to HRG

Series:	IN_VITRO DIFFERENTIATION SERIES
Species:	Human (Homo sapiens)
Genomic View:	Zenbu
Expression table:	FILE
Link to TET:	[{{{tet_file}}} TET]
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

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13048-139G6	MCF7 breast cancer cell line response to HRG	00hr15min	biol_rep1
13049-139G7	MCF7 breast cancer cell line response to HRG	00hr30min	biol_rep1
13050-139G8	MCF7 breast cancer cell line response to HRG	00hr45min	biol_rep1
13051-139G9	MCF7 breast cancer cell line response to HRG	01hr00min	biol_rep1
13052-139H1	MCF7 breast cancer cell line response to HRG	01hr20min	biol_rep1
13053-139H2	MCF7 breast cancer cell line response to HRG	01hr40min	biol_rep1
13054-139H3	MCF7 breast cancer cell line response to HRG	02hr00min	biol_rep1
13055-139H4	MCF7 breast cancer cell line response to HRG	02hr30min	biol_rep1
13056-139H5	MCF7 breast cancer cell line response to HRG	03hr00min	biol_rep1
13057-139H6	MCF7 breast cancer cell line response to HRG	03hr30min	biol_rep1
13058-139H7	MCF7 breast cancer cell line response to HRG	04hr	biol_rep1
13059-139H8	MCF7 breast cancer cell line response to HRG	05hr	biol_rep1
13060-139H9	MCF7 breast cancer cell line response to HRG	06hr	biol_rep1
13061-139I1	MCF7 breast cancer cell line response to HRG	07hr	biol_rep1
13062-139I2	MCF7 breast cancer cell line response to HRG	08hr	biol_rep1
13097-140D1	MCF7 breast cancer cell line response to EGF1	00hr00min	biol_rep2
13114-140E9	MCF7 breast cancer cell line response to HRG	00hr15min	biol_rep2
13115-140F1	MCF7 breast cancer cell line response to HRG	00hr30min	biol_rep2
13116-140F2	MCF7 breast cancer cell line response to HRG	00hr45min	biol_rep2
13117-140F3	MCF7 breast cancer cell line response to HRG	01hr00min	biol_rep2
13118-140F4	MCF7 breast cancer cell line response to HRG	01hr20min	biol_rep2
13119-140F5	MCF7 breast cancer cell line response to HRG	01hr40min	biol_rep2
13120-140F6	MCF7 breast cancer cell line response to HRG	02hr00min	biol_rep2
13121-140F7	MCF7 breast cancer cell line response to HRG	02hr30min	biol_rep2
13122-140F8	MCF7 breast cancer cell line response to HRG	03hr00min	biol_rep2
13123-140F9	MCF7 breast cancer cell line response to HRG	03hr30min	biol_rep2
13124-140G1	MCF7 breast cancer cell line response to HRG	04hr	biol_rep2
13125-140G2	MCF7 breast cancer cell line response to HRG	05hr	biol_rep2
13126-140G3	MCF7 breast cancer cell line response to HRG	06hr	biol_rep2
13127-140G4	MCF7 breast cancer cell line response to HRG	07hr	biol_rep2
13128-140G5	MCF7 breast cancer cell line response to HRG	08hr	biol_rep2
13163-141B4	MCF7 breast cancer cell line response to EGF1	00hr00min	biol_rep3
13180-141D3	MCF7 breast cancer cell line response to HRG	00hr15min	biol_rep3
13181-141D4	MCF7 breast cancer cell line response to HRG	00hr30min	biol_rep3
13182-141D5	MCF7 breast cancer cell line response to HRG	00hr45min	biol_rep3
13183-141D6	MCF7 breast cancer cell line response to HRG	01hr00min	biol_rep3
13184-141D7	MCF7 breast cancer cell line response to HRG	01hr20min	biol_rep3
13185-141D8	MCF7 breast cancer cell line response to HRG	01hr40min	biol_rep3
13186-141D9	MCF7 breast cancer cell line response to HRG	02hr00min	biol_rep3
13187-141E1	MCF7 breast cancer cell line response to HRG	02hr30min	biol_rep3
13188-141E2	MCF7 breast cancer cell line response to HRG	03hr00min	biol_rep3
13189-141E3	MCF7 breast cancer cell line response to HRG	03hr30min	biol_rep3
13190-141E4	MCF7 breast cancer cell line response to HRG	04hr	biol_rep3
13191-141E5	MCF7 breast cancer cell line response to HRG	05hr	biol_rep3
13192-141E6	MCF7 breast cancer cell line response to HRG	06hr	biol_rep3
13193-141E7	MCF7 breast cancer cell line response to HRG	07hr	biol_rep3
13194-141E8	MCF7 breast cancer cell line response to HRG	08hr	biol_rep3