Dissecting the transcriptional networks underlying breast cancer: NR4A1 reduces the migration of normal and breast cancer cell lines

Alexopoulou, Annika N., Leao, Maria, Caballero, Otavia L., Da Silva, Leonard, Reid, Lynne, Lakhani, Sunil R., Simpson, Andrew J., Marshall, John F., Neville, A. Munro and Jat, Parmjit S. (2010) Dissecting the transcriptional networks underlying breast cancer: NR4A1 reduces the migration of normal and breast cancer cell lines. Breast Cancer Research, 12 4: R51-1-R51-17. doi:10.1186/bcr2610

Author Alexopoulou, Annika N.
Leao, Maria
Caballero, Otavia L.
Da Silva, Leonard
Reid, Lynne
Lakhani, Sunil R.
Simpson, Andrew J.
Marshall, John F.
Neville, A. Munro
Jat, Parmjit S.
Title Dissecting the transcriptional networks underlying breast cancer: NR4A1 reduces the migration of normal and breast cancer cell lines
Journal name Breast Cancer Research   Check publisher's open access policy
ISSN 1465-5411
Publication date 2010-06-19
Sub-type Article (original research)
DOI 10.1186/bcr2610
Open Access Status DOI
Volume 12
Issue 4
Start page R51-1
End page R51-17
Total pages 17
Place of publication London, United Kingdom
Publisher Current Medicine Group
Language eng
Abstract Cell lines are extremely useful tools in breast cancer research. Their key benefits include a high degree of control over experimental variables and reproducibility. However, the advantages must be balanced against the limitations of modelling such a complex disease in vitro. Informed selection of cell line(s) for a given experiment now requires essential knowledge about molecular and phenotypic context in the culture dish.
Formatted abstract
Introduction: Breast cancer currently accounts for more than one-quarter of all female cancers and, despite the
great progress in treatment observed in the past few years, the need for identification of new gene targets that
can be used for diagnosis, prognosis and therapy is evident. A previous study identified the transcription factor
NR4A1 as a gene upregulated in primary breast cancer compared with normal tissue by microarray analysis and
sequencing technologies. The purpose of the study was to identify the role of NR4A1 in normal mammary
epithelial and breast cancer cell biology.
Methods: NR4A1 expression in breast tumours was assessed by semiquantitative and real-time PCR using RNA
from normal and tumour samples or breast cancer cell lines. Immunohistochemistry on tissue microarrays was
performed to check NR4A1 protein expression in breast tumours. MCF-10A and 226L normal mammary epithelial
cells as well as the tumour lines PMC42, ZR-75-1 and MDA-MB-231 were transduced with full-length NR4A1, and
the ability of NR4A1-overexpressing cells to migrate was tested using scratch wound or transwell migration assays.
Proliferation was measured using the MTT and BrdU assays, while apoptosis was determined by the Annexin V
assay. The ability of the cells to adhere to extracellular matrix was tested by adhesion assays and integrin cell
surface expression was measured by flow cytometry. Activation of the FAK as well as ERK1/2 and PI3K pathways
was checked by western blotting.
Results: Breast tissue microarray analysis showed NR4A1 expression in primary tumours, which was reduced in
higher grade and metastatic tumours. Ectopic expression of NR4A1 in MCF-10A, 226L, PMC42 and ZR-75-1 cells led
to reduced ability of the cells to migrate, while no differences were observed in their proliferation and apoptotic
index. NR4A1 expression altered the ability of the MCF-10A cells to adhere to the extracellular matrix and affected
cell surface expression of integrins.
Conclusions: NR4A1 acts as an antimigratory factor in two normal mammary epithelial and two breast cancer cell
lines tested. It is therefore possible that NR4A1 acts as an antimigratory factor in breast tumours, and further
studies should be conducted to understand the mechanisms involved.  © 2010 Alexopoulou et al.; licensee BioMed Central Ltd.
Keyword Orphan receptor TR3
Nuclear Receptors
Early genes
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Article# R51, pp.1-17

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
Official 2011 Collection
School of Medicine Publications
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Citation counts: TR Web of Science Citation Count  Cited 30 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 28 times in Scopus Article | Citations
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Created: Sun, 30 Jan 2011, 10:06:41 EST