Background
Methods
Patient samples
Characteristics | All patients (n = 85) |
---|---|
Median age (years) | 62 |
TNM classification: | |
pT0 | 11 |
pT1 | 0 |
pT2a | 10 |
pT2b | 30 |
pT3a | 28 |
pT3b | 17 |
T3N1M1b | 3 |
T3N1M1c | 2 |
Gleason grading: | |
5 | 4 |
6 | 8 |
7 | 53 |
8 | 15 |
9 | 5 |
Median follow-up (months) | 60 |
Metastatic relapse (n) | 4 |
Death (n) | 3 |
LuCaP 23.1 xenograft model
RNA extraction
DNA microarrays preparation and analysis
Tissue microarray (TMA) construction
Immunohistochemical analysis
Protein | Origin | Clone | Dilution | Antigen Retrieval |
---|---|---|---|---|
FGFR1 | SantaCruz | C15 | 1/200 | Demask (DAKO) |
MART1 | DAKO | A103 | 1/2 | Demask (DAKO) |
TACC1 | Upstate Biotechn. | 07–229 | 1/500 | Demask (DAKO) |
WT1 | Oncogene | WLM04 | 1/100 | Citrate (DAKO) |
PSA | DAKO | N1517 | rtu* | * |
RT-PCR analysis
TACC1 | F: AAATACGAAGAGACCCGGC |
R: TGTCCAGTTTCTCTTCTGCG | |
WT1 | F: ATGAGGATCCCATGGGCCAGCA |
R: CCTGGGACACTGAACGGTCCCCGA | |
MART1 | F: TGACCCTACAAGATGCCAAG |
R: TCAGCATGTCTCAGGTGTCT | |
Actin | F: AGCAAGAGAGGCATCCTGACC |
R: CTTCATGATGGAGTTGAAGGTAG |
GENE | N° of cycles | Denaturation | Annealing | Extension |
---|---|---|---|---|
WT1 | 35 | 94°C/30 sec | 64°C/30 sec | 72°C/30 sec |
MART1 | 30 | 94°C/1 min | 60°C/1 min | 72°C/2 min |
TACC1 | 30 | 94°C/1 min | 55°C/1 min | 72°C/2 min |
ACTIN | 25 | 94°C/30 sec | 55°C/30 sec | 72°C/30 sec |
Results
Overexpression of FGFR1, MART1, TACC1, and WT1 mRNAs in tumor progression in the LuCaP 23.1 carcinoma model
HR vs LUCAP 23.1 | Description |
---|---|
FGFR1
| Fibroblast growth factor receptor 1 |
KRT19
| Cytokeratin 19 |
CDK4
| Cyclin-dependent kinase 4 |
AMFR
| Autocrine motility factor receptor |
VEGFR1 | Vegf receptor 1/FLT1 |
ITGA3 | Integrin alpha 3/CD49C |
MUC1 | Polymorphic epithelial mucin |
GRB7 | GRB7 adaptor |
RELA | NFKb, p65 subunit |
BCL3 | BCL3 protein |
SRF | Serum response factor |
MAX | MAX transcription regulator |
NEO1 | Neogenin homolog 1 |
ITGB5
| Integrin beta 5 |
RXRA | Retinoic X receptor alpha |
IGFBP4 | Insulin-like growth factor binding protein |
CASP9 | Caspase 9 |
CREBBP | CREB binding protein |
MMP15 | Membrane-type matrix metalloproteinase 2 |
SMAD9 | MAD homolog 9 |
BAK | Apoptosis regulator bak |
MMP9 | 92 kDa gelatinase, matrix metalloproteinase-9 |
GATA1 | GATA-binding protein 1 |
MRP5 | ATP-binding cassette, sub-family C, member 5 |
PPP2R2C | PP2A BR gamma |
EPOR | Erythropoietin receptor |
FZD5 | Frizzled 5, WNT pathway |
HS vs LuCaP 23.1
| |
MART1 | Melan-a protein |
CSTB | Cystatin b |
NR1H3 | Nuclear receptor liver X receptor |
VINC | Vinculin |
STMN1 | Stathmin, phosphoprotein p19 |
STAT1 | signal transducer and activator of transcription 1 |
CBLB | Cas-Br-M ectropic retroviral transforming sequence b |
WNT2 | WNT factor 2 |
CDH12 | Cadherin 12 |
FGFR1, TACC1 and WT1 proteins display high levels of expression in advanced stages of human prostate carcinoma
Factors | benign | pT2 | pT3 |
p-value |
---|---|---|---|---|
No. of patients (%) | ||||
Age, years
| 1 | |||
<=50
| 0 | 0 | 0 | |
>50
| 11 | 39 | 46 | |
Gleason
|
0,057
| |||
5
| 0 | 3 | 1 | |
6
| 0 | 7 | 4 | |
7
| 0 | 25 | 28 | |
8
| 0 | 3 | 10 | |
9
| 0 | 0 | 4 | |
FGFR1 status
|
0.001
| |||
negative
| 9 | 27 | 17 | |
positive
| 2 | 12 | 29 | |
MART1 status
|
0.029
| |||
negative
| 11 | 30 | 43 | |
positive
| 0 | 9 | 3 | |
TACC1 status
|
0.756
| |||
negative
| 3 | 13 | 12 | |
positive
| 8 | 26 | 34 | |
WT1 status
|
0.008
| |||
negative
| 9 | 24 | 17 | |
positive
| 2 | 15 | 29 |