Background
Results
Retrospective clinical - histological data analysis
SURVIVAL GROUP | ARRAY | SURVIVAL TIME (days) | AGE (Year, Month) | SEX | BREED | TUMOR LOCATION | HISTOLOGY SUBTYPE | HISTOLOGY GRADE | POST OPERATIVE CHEMOTHERAPY |
---|---|---|---|---|---|---|---|---|---|
SHORT | 1 | 36 | 9,2 | F | Boxer | Rib | OB/CB/FB | highly malignant | No |
SHORT | 2 | 79 | 8,8 | M | Vizsla | distal radius | OB | highly malignant | Yes |
SHORT | 3 | 13 | 2,6 | F | Labrador retriever | proximal humerus | OB | highly malignant | No |
SHORT | 4 | 169 | 4,8 | M | Great Dane | distal radius | OB/FB | highly malignant | Yes |
SHORT | 5 | 121 | 7,9 | M | Rottweiler | proximal humerus | OB/CB/FB | medium malignant | Yes |
SHORT | 6 | 66 | 9,8 | M | Dobermann | ulna | OB/TL | medium malignant | No |
SHORT | 7 | 119 | 7,1 | F | Rottweiler | distal femur | OB | highly malignant | Yes |
SHORT | 8 | 66 | 11,6 | F | Belgian Shepherd (tervuren) | proximal humerus | OB | highly malignant | Yes |
SHORT | 9 | 77 | 7,0 | M | Great Dane | distal tibia | OB/FB | highly malignant | Yes |
SHORT | 10 | 164 | 6,7 | M | Dobermann | distal tibia | CB | highly malignant | No |
SHORT | 11 | 140 | 7,4 | M | Rottweiler | distal radius & ulna | OB/FB | highly malignant | No |
SHORT | 12 | 61 | 5,8 | F | Great Dane | distal femur | OB | medium malignant | Yes |
SHORT | 13 | 50 | 11,9 | M | Belgian Shepherd (tervuren) | distal tibia & fibula | OB | highly malignant | No |
SHORT | 14 | 47 | 6,8 | F | Rhodesian ridgeback | distal femur | OB/FB | highly malignant | Yes |
SHORT | 15 | 65 | 6,2 | M | Great Dane | proximal humerus | OB/CB | highly malignant | Yes |
SHORT | 16 | 6 | 6,1 | F | Rottweiler | mandible | OB/FB | highly malignant | No |
LONG | 17 | 352 | 9,8 | M | Dobermann | proximal tibia | OB/FB | medium malignant | Yes |
LONG | 18 | 274 | 8,8 | M | cross | ulna | OB/FB/TL | medium malignant | No |
LONG | 19 | 1752 | 3,8 | M | Stabyhoun | extraskeletal | OB/FB | highly malignant | Yes |
LONG | 20 | 307 | 4,3 | M | Great Dane | distal radius | CB | medium malignant | Yes |
LONG | 21 | 1619 | 3,7 | F | Siberian husky | mandible | OB/TL | highly malignant | No |
LONG | 22 | 1185 | 2,3 | F | Flatcoat retriever | rib | OB/TL | highly malignant | No |
LONG | 23 | 1289 | 5,0 | F | Rottweiler | distal radius | OB/FB | highly malignant | Yes |
LONG | 24 | 257 | 8,4 | F | Bouvier | distal radius | OB/FB | highly malignant | Yes |
LONG | 25 | 445 | 10,4 | M | Scottish collie | distal radius | OB | highly malignant | Yes |
LONG | 26 | 284 | 7,0 | F | cross | distal radius | OB/CB | highly malignant | Yes |
LONG | 27 | 239 | 8,4 | M | Rottweiler | scapula | OB/FB | highly malignant | Yes |
LONG | 28 | 705 | 8,3 | F | cross | metatarsus | FB | low malignant | Yes |
LONG | 29 | 348 | 9,0 | F | Mastiff | distal radius | OB | highly malignant | Yes |
LONG | 30 | 312 | 5,8 | M | Mastiff | scapula | OB/FB | highly malignant | Yes |
LONG | 31 | 920 | 7,3 | M | Belgian Shepherd (malinois) | mandible | OB/FB/CB | highly malignant | No |
LONG | 32 | 312 | 10,2 | M | Rottweiler | distal femur | OB/CB | highly malignant | Yes |
Parameter | Short survivors | Long survivors | (Fisher's exact test) P value |
---|---|---|---|
n
|
n
| ||
Gender
| > 0.999 | ||
Male | 9 | 9 | |
Female | 7 | 7 | |
Neuter status
| > 0.999 | ||
Neutered | 5 | 6 | |
Intact | 11 | 10 | |
Metastasis present at time of death
|
0.018**
| ||
Yes | 16 | 10 | |
No | 0 | 6 | |
Postoperative chemotherapy
| 0.458 | ||
No | 7 | 4 | |
Yes | 9 | 12 | |
Histological grade
| > 0.999 | ||
Low and medium | 3 | 4 | |
High | 13 | 12 |
Microarray data analysis
Gene Clone ID | Q-value (%) | Fold change | Gene description | Gene symbol |
---|---|---|---|---|
DG2-21g13 | 0.00 | 4.1 | WD repeat and SOCS box containing protein 2 | WSB2 |
DG2-23c15 | 0.00 | 3.3 | cofilin 2 | CFL2 |
DG2-72g4 | 0.00 | 2.7 | ankyrin repeat domain protein 17 isoform a | ANKRD17 |
DG42-128j23 | 0.00 | 2.1 | paraoxonase 1 | PON1 |
DG2-112n11 | 0.00 | 2.1 | Kinesin heavy chain (Ubiquitous kinesin heavy chain) | UKHC |
DG32-161c11 | 0.00 | 2.0 | WNK lysine deficient protein kinase 1 | WNK1 |
DG2-63l7 | 0.00 | 1.7 | nuclear receptor co-repressor 1 | NCOR1 |
DG32-237k11 | 0.00 | 1.6 | Ribosomal L1 domain containing protein 1(PBK1) | RSL1D1 |
DG11-239n21 | 0.00 | 1.6 | cell-cycle and apoptosis regulatory protein 1 | CCAR1 |
DG2-28n13 | 0.00 | 1.5 | 28S ribosomal protein S31, mitochondrial precursor | MRPS31 |
DG2-59p21 | 0.00 | 1.4 | #NA | |
DG2-90b10 | 4.61 | 2.2 | Heat shock protein HSP 90-alpha | HSP90 |
DG2-123a3 | 4.61 | 2.0 | Microsomal glutathione S-transferase 1 | MGST1 |
DG2-86b3 | 4.61 | 1.6 | Canis familiaris similar to T06D8.1a | |
DG8-102i3 | 4.61 | 1.5 | #NA | |
DG2-100e22 | 4.61 | 1.5 | COMM domain containing protein 8 | COMMD8 |
DG2-106f2 | 4.61 | 1.4 | Translocation protein SEC63 homolog | SEC63 |
DG2-130m14 | 4.61 | 1.4 | Canis familiaris similar to CG1218-PA | |
DG2-90c16 | 6.15 | 2.6 | #NA | |
DG43-1a15 | 6.15 | 2.4 | #NA | |
DG14-71c7 | 6.15 | 1.8 | serine/arginine repetitive matrix 1 | SRRM1 |
DG2-72p3 | 6.15 | 1.6 | Vacuolar ATP synthase subunit C | V-ATPase C |
DG32-216j13 | 6.15 | 1.6 | Flavin reductase (NADPH-dependent diaphorase) (FLR) | BVRB |
DG2-94j4 | 6.15 | 1.5 | Canis lupus familiaris high-mobility group box 1 | HMGB1 |
DG2-42j4 | 6.15 | 1.4 | FRA10AC1 protein isoform FRA10AC1-1) | |
DG42-89n19 | 6.15 | 1.4 | #NA | |
DG2-19i16 | 9.70 | 7.2 | #NA | |
DG2-18m22 | 9.70 | 4.6 | plasma glutamate carboxypeptidase | PGCP |
DG9-134g22 | 9.70 | 1.9 | ankyrin repeat domain 11 | ANKRD11 |
DG2-123m9 | 9.70 | 1.9 | CG12795-PA | |
DG14-14i19 | 9.70 | 1.7 | Stress-70 protein, mitochondrial precursor (HSAP9) (Mortalin) | MOT |
DG9-212f9 | 9.70 | 1.5 | CG12795-PA | |
DG2-60f11 | 9.70 | 1.5 | #NA | |
DG2-25k5 | 9.70 | 1.4 | SMC6 protein | SMC6 |
DG14-86l17 | 9.70 | 1.4 | #NA | |
DG2-24i24 | 9.70 | 1.4 | 60 kDa heat shock protein, mitochondrial precursor | HSP60 |
DG11-243e16 | 9.70 | 1.4 | splicing factor, arginine/serine-rich 2, interacting protein | SFRS2IP |
Quantitative real-time PCR analysis
Molecular functions and biological process analyses
Gene ID | Fold change | Gene description | Gene symbol |
---|---|---|---|
Metastasis-associated
| |||
DG2-23c15 | 3.3 | cofilin 2 | CFL2 |
DG14-14i19 | 1.7 | Stress-70 protein, mitochondrial precursor (HSAP9) (GRP 75) (Mortalin) | MOT |
DG2-72p3 | 1.6 | Vacuolar ATP synthase subunit C | V-ATPase C |
Drug resistance
| |||
DG2-112n11 | 2.1 | Kinesin heavy chain (Ubiquitous kinesin heavy chain) | UKHC |
DG2-123a3 | 2.0 | Microsomal glutathione S-transferase 1 | MGST1 |
DG2-72p3 | 1.6 | Vacuolar ATP synthase subunit C | V-ATPase C |
DG2-94j4 | 1.5 | high-mobility group box 1 | HMGB1 |
Cell cycle/proliferation
| |||
DG2-23c15 | 3.3 | cofilin 2 | CFL2 |
DG32-161c11 | 2.0 | WNK lysine deficient protein kinase 1 | WNK1 |
DG14-71c7 | 1.8 | serine/arginine repetitive matrix 1 | SRRM1 |
DG14-14i19 | 1.7 | Stress-70 protein, mitochondrial precursor (HSAP9) (GRP 75) (Mortalin) | MOT |
DG32-237k11 | 1.6 | Ribosomal L1 domain containing protein 1(PBK1 protein) | RSL1D1 |
DG11-239n21 | 1.6 | cell-cycle and apoptosis regulatory protein 1 | CCAR1 |
DG2-106f2 | 1.4 | Translocation protein SEC63 homolog | SEC63 |
DG2-25k5 | 1.4 | SMC6 protein | SMC6 |
Pathway analyses
Reference | Present study on canine osa | Srivastava et al., 2006 | Rajkumar, T. et al., 2008 | Srivastava et al., 2006 |
---|---|---|---|---|
Study approach
|
poor survival associated profile
|
high metastatic profile
|
drug resistant profile
|
osteogenic tumor profile
|
Samples used
|
32 primary tumors
|
human osa cell lines
|
cell line resistant to doxorubicin
|
10 tumors and 8 normal bones
|
Total differential expressed genes | 1426 | 1999 | 485 | 194 |
Total Mapped IDs on PANTHER® | 533 | 1185 | 295 | 159 |
Huntington disease | 12 | ▲ | ▲ | |
Wnt signaling pathway
|
10
| ▲ | ▲ | ▲ |
Chemokine/cytokine signaling pathway
|
8
| ▲ | ▲ | ▲ |
Integrin signalling pathway | 7 | ▲ | ▲ | |
Parkinson disease | 6 | ▲ | ▲ | |
Ubiquitin proteasome pathway | 6 | ▲ | ||
Alzheimer disease-presenilin pathway
|
5
| ▲ | ▲ | ▲ |
Cadherin signaling pathway | 5 | ▲ | ||
Endothelin signaling pathway | 5 | |||
Glycolysis | 5 | |||
Heterotrimeric G-protein signaling pathway | 4 | ▲ | ||
Angiogenesis | 3 | ▲ | ||
Cytoskeletal regulation by Rho GTPase | 3 | ▲ | ||
EGF receptor signaling pathway | 3 | ▲ | ||
FGF signaling pathway
|
3
| ▲ | ▲ | ▲ |
p53 pathway | 3 | ▲ | ||
PDGF signaling pathway
|
3
| ▲ | ▲ | ▲ |
T cell activation | 3 | ▲ | ▲ | |
TGF-beta signaling pathway | 3 | ▲ | ▲ | |
Androgen/estrogene/progesterone biosynthesis | 2 | |||
Apoptosis signaling pathway
|
2
| ▲ | ▲ | ▲ |
B cell activation | 2 | |||
Hedgehog signaling pathway | 2 | |||
Hypoxia response via HIF activation | 2 | |||
Interferon-gamma signaling pathway | 2 | |||
Interleukin signaling pathway
|
2
| ▲ | ▲ | ▲ |
Oxidative stress response | 2 | |||
p53 pathway feedback loops 2 | 2 | ▲ | ||
PI3 kinase pathway | 2 | ▲ | ▲ | |
Ras Pathway | 2 | ▲ | ▲ |
Discussion
Canine OS gene expression profiling as a model for human OS
Clinicopathological relevance for OS prognostic gene expression studies in dogs
Differentially expressed genes between short and long survivors: similarities with human OS
Pathway Analyses
Use of microarrays for OS prognosis prediction
Conclusion
Methods
Patient and tumor data
Statistical and survival analysis
RNA isolation and amplification
Microarray hybridization and data normalization
Microarray data analysis
Quantitative real-time PCR
Gene symbol | Forward and Reverse Primers | Temp (°C) |
---|---|---|
ANKRD17 | FW: 5'-AAGTAGCGCACCACCTTCAC-3' | 60.0 |
RW: 5'-CTAGCAGCAAATGGTGGACA-3' | ||
MRPS31 | FW: 5'-GAATTGGTCCTTGCTTTGGA-3' | 60.0 |
RW: 5'-ATCCAGTGGACGAAAGATGG-3' | ||
NCOR1 | FW: 5'-TCTTCCTCTGCGTTTTCCAT-3' | 59.6 |
RW: 5'-GCATCCCAAAAACTTTGGAC-3' | ||
MGST1 | FW: 5'-CGGACAGATGATAGGGTGG-3' | 62.0 |
RW: 5'-GATTTGGCTGGGGAAGG-3' | ||
RPS19 | FW: 5'-CCTTCCTCAAAAAGTCTGGG-3' | 61.0 |
RW: 5'-GTTCTCATCGTAGGGAGCAAG-3' | ||
HPRT | FW: 5'-AGCTTGCTGGTGAAAAGGAC-3' | 56.0 |
RW: 5'-TTATAGTCAAGGGCATATCC-3' |