Download - Potential Role of Monocyte Chemotactic Protein 1 (MCP-1) in the Breast Cancer Microenvironment
Potential Role of Monocyte Potential Role of Monocyte Chemotactic Protein 1 (MCP-1) Chemotactic Protein 1 (MCP-1)
in the Breast Cancer in the Breast Cancer MicroenvironmentMicroenvironment
MC Hartmann, RM Dwyer, MP Boyle, SM Potter and MJ Kerin
Department of Surgery, National University of Ireland, Galway
Introduction
Breast cancer is the leading type of female cancer in women in Ireland
Despite advances in diagnosis and therapy, there are few options available for treatment of advanced disease
Further understanding of the mechanisms involved in cancer spread from the primary tumour site is important to develop novel therapies
The primary tumour microenvironment plays an important part in tumour progression
Breast tumour microenvironment
Neoplastic epithelial cells co-exist in carcinomas with a biologically complex stroma, composed of various types of stromal cells and extracellular matrix
Cross talk between epithelial and stromal cells is mediated by chemokine signalling
Monocyte Chemotactic Protein 1 (MCP-1)
Tumour stromal cells
Tumour epithelial cells
MCP-1/ CCL2
Tumour stromal cells
Tumour epithelial cells
MCP-1/ CCL2
MCP-1 In Breast Cancer:MCP-1 In Breast Cancer: MCP-1 is a chemotactic cytokine, primary role in MCP-1 is a chemotactic cytokine, primary role in
inflammation. inflammation.
Whole tumour explants secrete high levels of MCP-1.Whole tumour explants secrete high levels of MCP-1.
Tumour stromal cells secrete significantly higher levels than Tumour stromal cells secrete significantly higher levels than normal stromal cells. normal stromal cells. Epithelial cells express its receptor, Epithelial cells express its receptor, CCR2CCR2
Effect of MCP-1 on epithelial cell gene expression is unknownEffect of MCP-1 on epithelial cell gene expression is unknown
Aim
Investigate the effect of Monocyte Investigate the effect of Monocyte Chemotactic Protein-1 on breast cancer Chemotactic Protein-1 on breast cancer cell cell proliferationproliferation and gene expression and gene expression
Methods
Breast cancer cell lines SK-BR-3
PR-, ER-, HER2/neu ++ T47 D
ER+, PR+, HER2/neu low MDA-MB 231
ER-, PR-, HER2/neu – Breast cancer cell lines incubated
with recombinant human MCP-1 for 72hrs Cells harvested and RNA extracted Changes in proliferation analysed
T47D
Cells incubated with MCP-1
Real Time Quantitative
PCR
ELK-1, VIL-2 MKi67, BAG-1, Bcl-2
PPIA, MRPL19
RNA Extraction
Methods
cDNA synthesis
Oxyluciferin+AMP + PPi + CO2
ATP+Luciferin+O2
Luciferase
Mg++ LIGHT
Cell Proliferation
Gene expression
MCP-1 mediated changes in gene expression in T47D
cells
0
1
2
3
4
5
6
BC L2 BAG1 VIL -2 E LK -1
Fol
d Cha
nge
in G
ene
Exp
ress
ion
* p<0.05
*
*
MCP-1 mediated changes in gene expression in SK-BR-3
cells
0
1
2
3
4
BCL2 BAG1 VIL-2 ELK-1
Fold
Cha
nge
in G
ene E
xpre
ssio
n
* p<0.05
**
MCP-1 mediated change in gene expression of MDA-
MB-231 cells
0
1
2
3
4
BCL2 BAG1 VIL-2 ELK-1
Fol
d Cha
nge
in G
ene
Exp
ress
ion
* p<0.05
*
Effect of MCP-1 on Cell Proliferation
Rela
tive
Pro
life
rati
on
(%
)
0
10
20
30
40
50
60
70
80
90
100
T47D T47D MDA-MB-231MDA-MB-231
+ MCP-1
Well Content
+ MCP-1
SK-BR-3 SK-BR-3
+ MCP-1
Summary
Up regulation of BAG-1 and Bcl-2 in Up regulation of BAG-1 and Bcl-2 in T47D and SK-BR-3 cellsT47D and SK-BR-3 cells
Up regulation of VIL 2 in MDA-MB-231Up regulation of VIL 2 in MDA-MB-231
No significant change in breast cancer No significant change in breast cancer cell proliferation in response to MCP-1cell proliferation in response to MCP-1
Conclusion
MCP-1 has a potentially important role MCP-1 has a potentially important role in the breast tumour microenvironment. in the breast tumour microenvironment.
The results presented suggest it may The results presented suggest it may exert its effect by promotion of tumour exert its effect by promotion of tumour cell survival through upregulation of cell survival through upregulation of anti-apoptotic factors and pro-anti-apoptotic factors and pro-migratory factors and thus may be a migratory factors and thus may be a target for therapeutic interventiontarget for therapeutic intervention..