Neoplasms are the second leading cause of death in the world after cardiovascular diseases. According to the World Health Organisation, 8.8 million people died of tumours in 2015 (
In canine and feline populations, the number of neoplasm cases also continues to increase around the world (60). According to Bronson (10) the death rate from tumours is the highest in older animals and reaches 45% in dogs older than 10 years. In cats, tumours are also most prevalent in older animals with an estimated death rate of 32% in cats older than 10 years. This can be attributed to the fact that companion animals live in the human environment and their predisposing risk factors are the same as humans. Advanced diagnostic techniques have also increased the detection rate for tumours in veterinary medicine (60).
In research centres, attempts are being made to identify new biomarkers that speed up and improve the quality of oncological diagnostics and have prognostic and predictive relevance in humans and animals with tumours (36). Rapid advances in knowledge of biochemical and especially molecular mechanisms responsible for oncogenesis and the progression of various types of tumours facilitate the development of new, more effective diagnostic methods and therapeutic protocols. The existing methods should be integrated, and the results of clinical evaluations, post-mortem examinations, histopathological, immunohistochemical, cytological, and molecular analyses should be used to deepen our understanding of tumour development and to improve diagnosis and treatment. The development of reliable parameters with prognostic and predictive applicability poses one of the greatest challenges in the treatment of tumours. Various biological, clinical, histological, immunohistochemical, and molecular parameters have been evaluated to date, but the choice of the most effective markers remains an open issue (28, 36, 46).
Cyclooxygenase-2 (COX-2) is an important diagnostic and prognostic biomarker to which human oncology has increasing recourse, while veterinary oncology does so to a lesser extent (50). COX-2 is not yet used as a biomarker in routine cancer screening in human medicine (36). This enzyme is overexpressed in humans presenting with various types of tumours and in selected types of tumours in animals, particularly in dogs. These findings indicate that COX-2 could be a potentially effective biomarker with diagnostic, therapeutic, prognostic, and predictive relevance in oncology. However, further research is required, particularly in veterinary medicine, because researchers are divided in their opinions on the expression of COX-2 in different types of tumours (7, 41).
Cyclooxygenase (COX) is an enzyme belonging to the myeloperoxidase family, which catalyses conversion of the arachidonic acid to prostanoids. These comprise prostaglandins, prostacyclin and thromboxane, which are bioactive proteins which regulate various physiological processes in human and animal organisms (14). COX-2 as an inducible isoform is found at low levels in mammalian cells and it is not generally detected in physiological conditions, although the presence of COX-2 was observed in the central nervous system, alimentary tract, heart, kidney, eye, and reproductive organs. In the beginning COX-2 was connected with the response to stress, but its overexpression is also found during fever, pain, and inflammation (50). In several studies, increased COX-2 expression was also demonstrated in neoplastic tissues, which suggests participation of this enzyme in carcinogenesis. COX-2 overexpression causes the cells’ phenotype to change from benign to malignant, which is connected with disruption of their growth and proliferation and an increase in cells’ ability to evade apoptosis and immune response, promote new blood vessels, and raise their invasive potential (50). More details of the function of COX-2 in the organism and the significance of COX-2 in oncogenesis were presented in our previous paper (57).
The mechanisms of COX-2 participation in oncogenesis are very complicated and weakly understood, particularly in animals. They comprise interactions between tumour cells and the surrounding microenvironment to create the best conditions for their growth, proliferation, and dissemination (22). A strict connection between chronic inflammatory processes and carcinogenesis was observed, when overexpression of COX-2 can contribute to the conversion of inflammation into cancer. Gandhi
Higher COX-2 expression was found in tumours of various organs in humans (17, 28) and dogs (19, 39), and to a lesser extent in cats (7, 39), which is often connected with a higher histological grade of tumour malignancy, worse prognosis, and shortening of overall survival (OS) (13, 23). In Poland previous studies evaluating COX-2 expression in animal tumours are relatively limited, comprising mainly canine mammary (44) and mast cell tumours (27).
Studies of COX-2 expression in animal populations in various countries have demonstrated that it is overexpressed in various canine epithelial tumours, including adenocarcinomas and carcinomas of the mammary gland, ovarian and prostate tumours, transitional cell (urothelial) carcinoma (TCC), colorectal and small intestine tumours, squamous cell carcinoma (SCC) of the skin and oral cavity, osteosarcoma, and melanoma (19, 20, 39, 46). The enzyme is minimally expressed or is not expressed in canine lymphoma, sarcoma, fibrosarcoma, glioma, and mastocytoma (41).
Research conducted on small populations of feline patients has revealed that unlike in dogs, COX-2 is expressed more weakly and far less frequently in cats. The enzyme was found in one study by Millanta
Several research works on COX-2 expression in canine and feline neoplasms have been written in recent years. In most of them, overexpression of COX-2 in tumours occurring in dogs and cats was found. Millanta
Some studies concerned cutaneous and oral SCC in dogs and cats. Millanta
Other canine and feline neoplasms were also the object of studies. Gregorio
The recent very considerable progress in molecular techniques for tumour diagnosis, including the emergence of DNA and RNA sequencing tools, single-nucleotide-polymorphism–based genotyping, and evaluation of mRNA, ncRNA, and miRNA profiles, as well as the achievements in proteomics, metabolomics, and bioinformatics have expanded our understanding of the molecular mechanisms responsible for tumour initiation, progression, and response to treatment, in particular in combination with histopathological and clinical evaluations (28, 36, 50). The results of intensive large-population studies are rapidly implemented in tumour treatment, albeit with varying degrees of success. However, they are less often used to develop sensitive and specific markers with diagnostic, prognostic, and predictive relevance in cancer medicine. Despite the subordinacy of this goal, the number of identified biomarkers with potential use in cancer diagnosis and treatment as well as prognostic and predictive biomarkers continues to increase (36).
Prognostic biomarkers such as β-tubulin, carbohydrate antigen 19-9 (CA19-9), cell-surface antigen CD44 (CD44), carcinoembryonic antigen, ColoPrint, circulating tumour cell, cyclin D1, E-cadherin, epidermal growth factor receptor (EGFR), inhibitor of growth protein 3 (ING3), Ki-67, matrix metalloproteinase-2, p-21, retinoblastoma gene, ribonucleotide reductase M1, and vascular endothelial growth factor (VEGF) are used to evaluate the malignant potential of tumours and measure the patients’ OS and progression-free survival (PFS) without treatment and after conventional treatment. These biomarkers are applied to qualify patients for treatment, but they do not support the prediction of treatment outcomes (Table 1) (36).
Prognostic biomarkers for survival in cancer medicine (36)
Prognostic biomarker | Type of cancer | Clinical significance | Detection | Clinical use |
---|---|---|---|---|
Beta-tubulin | NSCLC | High expression of β-tubulin confers worse prognosis | IHC | No |
BRCA1 | Breast | High expression of BRCA1 confers worse prognosis in untreated patients | IHC | Yes |
NSCLC | High expression of BRCA1 confers worse prognosis in untreated patients | qRT-PCR | No | |
CA19-9 | Pancreatic | Higher preoperative CA19-9 levels are associated with lower resectability, more advanced stage and inferior survival I | IHC | No |
CAIX | RCC | High expression of CAIX is associated with a better prognosis | IHC | No |
CD44 | Bladder | Expression of CD44 is associated with poor prognosis | qRT-PCR | No |
CEA | CRC | Elevated preoperative CEA levels in resectable colorectal cancer is associated with poor prognosis | IHC | Yes |
c-KIT | GIST | GIST patients have a better prognosis if they harbour a mutation in exon 11 of the c-KIT gene | Pathway detection via FDG-PET | Yes |
ColoPrint | CRC | Prognosis for colorectal cancer patients | Microarray | Yes |
CTC ( |
Melanoma | Increased number of circulating melanoma cells is associated with poor prognosis | Circulating tumour cells | No |
CRC | Colorectal patients with 3 CTC/7.5 ml of peripheral blood were associated with shorter PFS and OS, |
Circulating tumour cells | Yes | |
Breast | Breast cancer patients with 5 CTC/7.5 mL of peripheral blood are associated with shorter PFS and OS, |
Circulating tumour cells | Yes | |
Prostate | Circulating tumour cells | Yes | ||
Cyclin D1 | Bladder | Expression of Cyclin D1 is associated with low grade, low stage and recurrence | IHC | No |
Cyclin E | Bladder | Expression of Cyclin E is associated with low stage and survival | IHC | No |
E-Cadherin | Bladder | E-Cadherin is associated with poor prognosis | IHC | No |
EGFR | Bladder | Overexpression of EGFR is associated with high grade and high stage | IHC | No |
NSCLC | High gene copy number of EGFR in NSCLC patients is associated with poor prognosis | FISH / SA | No | |
EGFR mutation in NSCLC patients is associated with better prognosis in untreated patients | ||||
Rectal | Overexpression of EGFR in rectal cancers is also associated with poor prognosis | IHC | No | |
ER | Breast | Patients with ER-positive breast tumours have better survival than patients with hormonal negative tumours | IHC | Yes |
eXageneBC | Breast | Provides prognosis in node-positive or node-negative breast cancer patients | FISH | Yes |
Her2/neu | Breast | Patients with Her2/neu-positive breast tumours are more aggressive and have a worse prognosis compared to Her2/neu-negative tumours | FISH | Yes |
Bladder | Overexpression of Her2/neu is associated with high grade, high stage, poor survival and metastasis in bladder cancer | IHC | No | |
GIST | Overexpression of Her2/neu in advanced gastric cancer patients is associated with poor prognosis | IHC | No | |
Her3 | Melanoma | Correlation with increased cell proliferation, tumour progression and reduced survival in melanoma patients | IHC | No |
ING3 | Melanoma | Reduced nuclear expression associated with poor disease- specific survival in melanoma patients | IHC | No |
ING4 | Melanoma | Reduced levels of ING4 in melanoma patients is associated with melanoma thickness, ulceration and poor disease-specific survival and overall survival | IHC | No |
Ki-67 | Bladder | Expression of Ki-67 is associated with progression and recurrence in bladder cancer | IHC | No |
Breast | Expression of Ki-67 is associated with proliferation and progression in breast cancer | IHC | No | |
K-ras | NSCLC | K-ras mutation is associated with poor prognosis in NSCLC patients | SA | Yes |
LOH at 18q | CRC | Associated with metastasis and poor prognosis in colorectal tumours | PCR | No |
MammaPrint | Breast | A 70-gene prognostic assay used to identify breast cancer cases at the extreme end of the spectrum of disease outcome by identifying patients with good or very poor prognosis | Microarray | Yes |
Mammostrat | Breast | This standard purely prognostic test uses five antibodies with manual slide scoring to divide cases of ER-positive, lymph node negative breast cancer tumours treated with tamoxifen alone into low-, moderate- or high-risk groups | IHC | Yes |
MMP-2 | Bladder | Expression of MMP-2 is associated with poor prognosis in bladder cancer patients | PCR | No |
MSI status | CRC | High frequency MSI colorectal tumours are associated with better prognosis and show improved relapse-free survival | IHC | No |
NCOA3 | Melanoma | Increased levels in melanoma patients correspond to poor relapse-free survival and disease-free survival | IHC | No |
Oncotype DX | Breast | A 21-gene multiplex test used for prognosis to determine 10- year disease recurrence for ER-positive, lymph node negative breast cancers using a continuous variable algorithm and assigning a tripartite recurrence score | qRT-PCR | Yes |
p21 | Bladder | Overexpression of p21 is associated with poor prognosis | IHC | No |
p53 | Bladder | Overexpression of p53 is associated with poor prognosis | IHC | No |
NSCLC | High expression of p53 in NSCLC patients confers worse prognosis in untreated patients | IHC | No | |
NSCLC | p53 mutation in NSCLC patients is associated with worse prognosis | SA | No | |
PR | Breast | Patients with PR-positive breast tumours have better survival than patients with hormonal-negative tumours | IHC | Yes |
Rb | Bladder | Overexpression of Rb is associated with poor prognosis | IHC | No |
RRMI | NSCLC | High expression of RRMI in NSCLC patients confers better prognosis in untreated patients | AQUA | No |
VEGF | RCC | Overexpression of VEGF is associated with poor prognosis in clear cell renal carcinoma patients | IHC | Yes |
AQUA – automated quantitative analysis; CA19-9 – carbohydrate antigen 19-9; CAIX – carbonic anhydrase IX; CEA – carcinoembryonic antigen; CRC – colorectal tumour; CTC – circulating tumour cells; EGFR – epidermal growth factor receptor; ER – oestrogen receptor; FDG – 18F-fluorodeoxyglucose; FISH – fluorescent in situ hybridisation; GIST – gastrointestinal stromal tumour; IHC – immunohistochemistry; LOH – loss of heterozygosity; MMP-2 – matrix metalloproteinase-2; MSI – microsatellite instability; NSCLC – non-small cell lung cancer; OS – overall survival; PET – Positron emission tomography; PFS – progression-free survival; PR – progesterone receptor; qRT-PCR – quantitative real time polymerase chain reaction; Rb b – retinoblastoma; RCC – renal cell carcinoma; RRMI – ribonucleotide reductase messenger 1; SA – sequence analysis; VEGF – vascular endothelial growth factor
Predictive biomarkers such as epidermal growth factor receptor 1 (EGFR1), excision repair cross-complementation group 1, O(6)-methylguanine-DNA methyltransferase, thymidyne phosphorylase, and phosphatase and tensin homolog support objective identification of individuals who are more likely to benefit from a given treatment or aid the evaluation of differences in the outcomes of two or more treatment procedures in view of their toxicity (Table 2) (36).
Predictive biomarkers for treatment selection in cancer medicine (36)
Predictive biomarker | Type of cancer | Clinical significance | Detection | Clinical use |
---|---|---|---|---|
BRCA1 | NSCLC | High expression of BRCA1 in NSCLC patients predicts resistance to cisplatin-based chemotherapy | qRT-PCR | No |
Breast | High expression of BRCA1 in breast cancer can predict response to chemotherapy | IHC | Yes | |
CAIX | RCC | Expression of CAIX in renal cell carcinoma is predictive of sensitivity of treatment with interleukin-2 therapy | IHC | No |
c-KIT | GIST | GIST patients carrying the mutation on exon 11 of the c-KIT gene benefit from imatinib and sunitinib treatment, however most patients develop resistance to these over time | SA | Yes |
EGFR1 | NSCLC | EGFR1 mutations in patients with NSCLC are predictive for response to either gefitinib or erlotinib treatment | IHC | Yes |
CRC | EGFR1 gene amplification appears to be a predictive factor for response to anti-EGFR1 antibody treatment in CRC | PCR | Yes | |
ER | Breast | High cellular expression of ER predicts benefit from tamoxifen- based chemotherapy | IHC | Yes |
ERCC1 | NSCLC | High expression of ERCC1 in NSCLC patients predicts resistance to cisplatin-based chemotherapy | IHC | No |
Her2/neu | Breast | Breast cancer patients with Her2/neu overexpressing tumors benefit from treatment with trastuzumab in the metastatic as well as in the adjuvant setting | FISH | Yes |
Gastric | Expression of Her-2/Neu in gastric cancer is predictive of patient sensitivity towards treatment with 5-FU, doxorubicin, trastuzumab and platinum-based chemotherapy | FISH | No | |
K-ras | NSCLC | K-ras mutation positivity in NSCLC patients predicts lack of benefit from adjuvant chemotherapy in early disease and resistance to treatment with EGFR TKI in advanced disease | SA | Yes |
CRC | K-ras mutation positivity in stage IV CRC patients predicts considerably less benefit from EGFR-specific antibody like cetuximab and panitumumab | PCR | Yes | |
LOH at 18q | CRC | Useful in identifying patients with resected stage III colon cancer most likely to benefit from 5-FU based adjuvant chemotherapy | PCR | No |
MGMT | Glioblastoma | Methylation of MGMT promoter is predictive of sensitivity of glioblastoma to temozolomide | PCR | No |
NuvoSelect | Breast | A combination of several pharmacogenomic genesets used primarily to guide selection of therapy in breast cancer patients. This test also provides the ER and HER2 mRNA status | Microarray | Yes |
p53 | NSCLC | High p53 expression in NSCLC patients predicts sensitivity to cisplatin-based chemotherapy, however p53 mutation is predictive of resistance to cisplatin-based chemotherapy | IHC/SA | No |
PR | Breast | High cellular expression of PR predicts benefit from tamoxifen- based chemotherapy | IHC | Yes |
Roche AmpliChip | Breast | Low expression of CYP2D6 predicts resistance to tamoxifen- based chemotherapy in breast cancer patients | Microarray | Yes |
Rotterdam Signature | Breast | A 76-gene assay used to predict recurrence in ER-positive breast cancer patients treated with tamoxifen | Microarray | Yes |
RRMI | NSCLC | High expression of RRM1 in NSCLC patients predict resistance to cisplatin-based chemotherapy | qRT-PCR | No |
TP | GIST | Predictive of sensitivity of treatment to 5-FU- and capcetabine- based chemotherapy in gastric cancer patients | IHC/PCR | No |
CRC | Expression of TP in metastatic colorectal patients is predictive of sensitivity of treatment to 5-FU and capcetabine based chemotherapy | IHC/qRT-PCR | No | |
PTEN | Breast | PTEN mutation can result in reduced sensitivity of treatment with trastuzumab in breast cancer patients | IHC | No |
CAIX – carbonic anhydrase IX; CRC – colorectal tumour; EGFR – epidermal growth factor receptor; ER – oestrogen receptor; ERCC1 – excision repair cross-complementation group 1; FISH – fluorescent
Some biomarkers, including breast cancer gene (BRCA1), carbonic anhydrase IX (CAIX), oestrogen receptor (ER), progesterone receptor (PR), tumour suppressor protein (p53), human epidermal growth factor receptor 2 (HER2/neu), and Kirsten rat sarcoma oncogene, have both prognostic and predictive relevance (36). Multigene panel tests are also used to identify groups of up to several dozen genes, mainly in the diagnosis of breast cancer, for which application MammaPrint (59) or Mammostrat (49) are examples of available assays.
The increased expression of COX-2 in various types of tumours, in particular in dogs, but also in cats, suggests possibilities for its utilisation in practice. Its introduction may be feasible into routine evaluation as a diagnostic, therapeutic, prognostic, and predictive biomarker in small-animal veterinary oncology especially, in like manner to how it is exploited to a certain extent in human oncology (5).
In human medicine, COX-2 overexpression in tumour patients is often associated with poor prognosis and reduced OS and/or PFS (30). The applicability of COX-2 in the diagnosis of canine tumours requires further research because the results of studies evaluating these associations and another between the overexpression and response to treatment are contradictory (19, 20). Correlations with poor prognosis and reduced OS have been observed in canine mammary gland carcinoma (47), whereas no such relationships have been reported in canine prostatic carcinoma (55). Queiroga
Because limited veterinary literature is available regarding prognostic biomarkers for canine renal cell carcinoma (CRCC), Carvalho
Experimental, clinical, and epidemiological studies have demonstrated that non-steroidal anti-inflammatory drugs (NSAIDs), in particular selective COX-2 inhibitors (coxibs), effectively inhibit tumour progression and improve chemotherapy outcomes in human patients (16). Specific COX-2 inhibitors, including celecoxib and rofecoxib, have been developed to minimise some mainly gastrointestinal side-effects of NSAIDs. In addition to anti-inflammatory, analgesic and antipyretic effects, these compounds also deliver anticarcinogenic effects by inhibiting the production of prostanoids. In some cases, however, anticarcinogenic effects were observed independently of COX-2 inhibition. Tamura
The discovery that coxibs possess anticarcinogenic properties laid the groundwork for clinical research in human oncology, which initially focused on coxibs’ chemopreventive and subsequently on its chemotherapeutic effects. Initial studies demonstrated that coxibs are effective in the treatment of familial adenomatous polyposis (FAP), but subsequent large-population research programmes revealed that coxibs have significant cardiovascular side-effects. Due to safety concerns, rofecoxib has been withdrawn from the pharmaceutical market, and celecoxib is presently prescribed only as a chemopreventive agent for FAP (3). However, a review of 72 research programmes carried out by Harris (24) did not confirm those concerns and found that coxibs caused side-effects only in patients with a higher risk of cardiovascular diseases.
The therapeutic effects of NSAIDs in cancer treatment have been confirmed by numerous studies which investigated the combined application of NSAIDs, radiotherapy, and chemotherapy in human patients (34). Overexpression of COX-2 has also been observed in some canine and feline tumours, and research findings indicate that this enzyme could be more widely used as a biomarker in veterinary medicine, in the diagnosis and treatment of cancer with the use of COX-2 inhibitors (39). This biomarker could be applied to identify patients where the use of non-selective and, in particular, selective COX-2 inhibitors could reduce COX-2 overexpression, limit tumour progression and increase survival rates (16, 34).
The use of NSAIDs in the treatment of canine and feline tumours has been investigated by relatively few studies, which, nevertheless, produced interesting results. Boria
Although there are several pieces of evidence supporting an important role of COX-2 in tumour development and progress in humans and animals, further studies are necessary to explore its significance. Subsequent investigation will elucidate the agency of COX-2 in oncogenesis, determine COX-2 expression levels in various types of canine and, in particular, feline tumours, assess the diagnostic, therapeutic, prognostic, and predictive relevance of this biomarker precisely, and evaluate the usefulness of NSAIDs in the chemoprevention and chemotherapy of canine and feline tumour patients.