Introduction
Methods
Cell culture and transfections
RNA amplification, labeling and hybridization to Agilent microarrays
Gene expression analysis
Bioinformatics analysis
Chemokine array
Immunohistocytochemistry
Immunofluorescence
Experimental metastasis assays
Antibody-mediated depletion experiments
CD11b+/Ly-6G+ cell isolation
RNA isolation and real-time RT-PCR
Statistical analysis
Results
Gene expression profiling reveals distinct expression patterns associated with 4T1 subpopulations derived from different metastatic sites
4T1-derived metastatic breast cancer populations possess distinct chemokine profiles
Ingenuity canonical pathways
|
p-value
|
Ratio
|
Molecules
|
---|---|---|---|
Granulocyte adhesion and diapedesis
| 4.94E-06 | 7.18E-02 | VCAM1,SDC1,MMP3,MMP10,CLDN7,CCL5,CXCL5,IL18RAP,CXCL10,ITGB2,MMP9,CLDN3,MMP19 |
Inhibition of matrix metalloproteases | 3.54E-05 | 1.50E-01 | SDC1,MMP3,RECK,MMP10,MMP9,MMP19 |
MMP19 hepatic fibrosis/hepatic stellate cell activation | 8.91E-05 | 6.45E-02 | 02VCAM1,CTGF,FGFR1,PDGFRA,LBP,CCL5,PDGFC,MMP9,PGF,IL18RAP |
Regulation of the epithelial-mesenchymal transition pathway | 1.81E-04 | 5.61E-02 | MAP2K6,CDH1,JAG2,WNT10B,ESRP2,FGFR1,PARD6G,JAG1,NOTCH1,MMP9,CLDN3 |
Agranulocyte adhesion and diapedesis
| 2.09E-04 | 5.76E-02 | CXCL10,ITGB2,VCAM1,MMP3,MMP10,CLDN7,CCL5,CXCL5,MMP9,CLDN3,MMP19 |
HIF1alpha signaling | 2.68E-04 | 7.14E-02 | MMP3,MMP10,PDGFC,LDHA,MMP9,LDHB,PGF,MMP19 |
Antigen presentation pathway
| 3.05E-04 | 1.19E-01 | B2M,PSMB9,HLA-B,PSMB8,TAP1 |
Bladder cancer signaling | 5.99E-04 | 7.22E-02 | CDH1,MMP3,MMP10,PDGFC,MMP9,PGF,MMP19 |
IL-8 signaling
| 9.59E-04 | 4.44E-02 | ITGB2,GNB4,CDH1,ANGPT2,VCAM1,CCND2,MYL12B,PDGFC,MMP9,PGF |
Leukocyte extravasation signaling
| 1.48E-03 | 4.76E-02 | ITGB2,VCAM1,EDIL3,MMP3,CD44,MMP10,CLDN7,MMP9,CLDN3,MMP19 |
VDR/RXR activation | 1.63E-03 | 6.82E-02 | CXCL10,SPP1,GADD45A,SEMA3B,VDR,CCL5 |
Pyruvate fermentation to lactate | 2.53E-03 | 2.22E-01 | LDHA,LDHB |
ILK signaling | 3.47E-03 | 4.39E-02 | MAP2K6,ITGB2,PARVB,CDH1,VIM,PDGFC,MMP9,DSP,PGF |
Pathogenesis of multiple sclerosis | 8.71E-03 | 2.00E-01 | CXCL10,CCL5 |
Sphingosine-1-phosphate signaling | 8.97E-03 | 4.88E-02 | PLCD1,CASP12,CASP1,PDGFRA,CASP4,PDGFC |
Axonal guidance signaling | 1.04E-02 | 2.90E-02 | WNT10B,BDNF,MMP10,ADAMTS9,PDGFC,SEMA4C,PGF,PLCD1,GNB4,MYL12B,BMP7,SEMA3B,MMP9,SEMA7A |
Atherosclerosis signaling | 1.39E-02 | 4.35E-02 | ITGB2,VCAM1,MMP3,S100A8,PDGFC,MMP9 |
Sorbitol degradation I | 1.62E-02 | 2.00E-01 | SORD |
Asparagine biosynthesis I | 1.62E-02 | 1.25E-01 | ASNS |
Alanine biosynthesis III | 1.62E-02 | 3.33E-01 | NFS1 |
Colorectal cancer metastasis signaling | 1.73E-02 | 3.36E-02 | GNB4,CDH1,WNT10B,MMP3,MMP10,PDGFC,MMP9,PGF,MMP19 |
p53 signaling | 1.75E-02 | 4.63E-02 | TP53INP1,CCND2,GADD45A,PLAGL1,SFN |
Role of IL-17A in psoriasis | 1.81E-02 | 1.43E-01 | S100A8,CXCL5 |
Chondroitin sulfate degradation (metazoa) | 2.09E-02 | 8.70E-02 | CD44,HEXA |
Human embryonic stem cell pluripotency | 2.19E-02 | 3.73E-02 | WNT10B,BDNF,FGFR1,PDGFRA,BMP7,PDGFC |
Notch signaling | 2.33E-02 | 6.98E-02 | JAG2,JAG1,NOTCH1 |
Dermatan sulfate degradation (metazoa) | 2.38E-02 | 8.70E-02 | CD44,HEXA |
Thyroid cancer signaling | 2.66E-02 | 6.82E-02 | CXCL10,CDH1,BDNF |
Alanine degradation III | 3.21E-02 | 1.67E-01 | GPT2 |
Alanine biosynthesis II | 3.21E-02 | 1.67E-01 | GPT2 |
Formaldehyde oxidation II (glutathione-dependent) | 3.21E-02 | 1.00E-01 | ESD |
Glutamine degradation I | 3.21E-02 | 2.00E-01 | GLS2 |
GADD45 signaling | 4.09E-02 | 8.33E-02 | CCND2,GADD45A |
T lymphocytes are recruited to breast cancer liver and lung metastases
A granulocytic infiltrate is recruited in breast cancer-derived lung and liver metastases
Gr-1+ cell depletion differentially affects the formation of bone, lung and liver metastases
Neutrophil recruitment persists at the lesion margins during progression of breast cancer-derived liver metastases
N2-polarized neutrophil recruitment increases at the lesion margins during progression of breast cancer-derived liver metastases
D7
| |||
---|---|---|---|
Location
|
0 < metastatic lesion area/liver area (%) <1.79
| ||
% N2 neutrophils
a
|
Error
|
P
value
| |
Distal | 60.84 | +/− 9.39 | |
Invasive front | 90.19 | +/− 6.04 | <0.00001 |
D14
| |||
Location
|
Low metastatic burden
| ||
2.16 < metastatic lesion area/liver area (%) <4.19
| |||
% N2 neutrophils
a
|
Error
|
P
value
| |
Distal | 79.58 | +/− 5.93 | |
Invasive front | 94.92 | +/− 3.56 | <0.0001 |
D14
| |||
Location
|
High metastatic burden
| ||
13.62 < metastatic lesion area/liver area (%) <54.12
| |||
% N2 neutrophils
a
|
Error
|
P
value
| |
Distal | 88.21 | +/− 3.53 | |
Invasive front | 93.63 | +/− 2.80 | 0.003 |