Feline Mammary Gland Research Articles

Distribution of Inflammatory Infiltrate in Feline Mammary Lesions: Relationship With Clinicopathological Features.

Inflammation is a frequent finding in feline mammary neoplasms. Recent research suggests that the presence and location of tumour-associated immune cells might play a significant role in the clinical outcome of feline mammary carcinomas. The present study aimed to characterise the overall inflammatory infiltrates in healthy, hyperplastic/dysplastic, benign and malignant lesions of the feline mammary gland, and to evaluate its association with clinicopathological features. Perilesional and intralesional inflammatory foci were evaluated in 307 lesions from 185 queens, and categorised according to its distribution and intensity. The presence, location and density of tertiary lymphoid structures were also assessed. A control group included 24 queens without mammary changes. The presence of intralesional and perilesional inflammatory infiltrate was observed in a majority of the lesions (80.8% and 90.2%, respectively), but differed according to the type of mammary lesion, being more remarkable in malignant neoplasms. Only scarce individual cells were observed in 28.1% of the normal mammary glands. Data analysis revealed statistically significant associations (p < 0.05) between the presence of a more prominent intralesional and perilesional inflammatory reaction and several clinicopathological features associated with worse prognosis, including clinical stage, tumour size, mitotic count, lymphovascular invasion and lymph node metastasis. Furthermore, tertiary lymphoid structures were significantly more frequent in tumours with an infiltrative growth and lymph node metastasis. According to our results, the inflammatory reaction present in different types of feline mammary lesions is associated with the development of more aggressive tumours.

Epidemiology and classification for canine and feline mammary gland tumors: a histopathological survey of 437 mammary gland tumor biopsies performed in a secondary care hospital in Chiang Mai, Thailand from 2012 to 2019.

Metastatic disease resulting from mammary gland tumors (MGTs) is a known cause of death among dogs and cats. Keys to successful prevention and management strategies involve the accurate recording of diagnostic data. This retrospective study reviewed the epidemiology and classification of canine mammary gland tumors (CMTs) and feline mammary gland tumors (FMTs), as well as the factors including sex, age, and breed related to the occurrence of these tumors. Accordingly, 1,736 tumor biopsy cases were reported from 2012 to 2019 at Chiang Mai University Small Animal Hospital, Thailand, with 1,639 canine tumor biopsy cases and 97 feline tumor biopsy cases. The proportion of CMTs was reported at 24.5% (401/1,639) for all canine tumor biopsy cases. Benign and malignant tumors were reported at 14.5% (58/401) and 85.5% (343/401) for all CMT cases, respectively. The mean age of dogs affected by benign CMTs was 9.0±3.0 years, which was significantly lower than for malignant CMTs at 9.9±2.8 years (P=0.0239). According to histopathological classification, benign mixed tumors and simple carcinoma types were highest among benign and malignant CMT cases, respectively. Moreover, female dogs were at significantly higher risk of developing mammary gland tumors (OR = 45.8, 95% CI [3.9-86.0], P<0.0001) than male dogs, as well as older dogs (>8 years) (OR = 1.7, 95% CI [1.2-2.2], P=0.0001) compared to young ones (≤8 years). The proportion of FMTs was 37.1% (36/97) for all feline tumor biopsy cases. Benign and malignant tumors for all FMTs were reported at 16.7% (6/36) and 83.3% (30/36), respectively. According to histopathological classifications, adenoma and simple carcinoma were present in the highest proportion among benign and malignant FMTs, respectively. Female cats were at a significantly higher risk of developing mammary gland tumors than male cats (OR=25.7, 95% CI [3.9-272.8], P<0.0001). There was a high proportion of MGT cases compared with other tumor cases reported in a secondary care hospital in Chiang Mai, Thailand from 2012 to 2019, and malignant tumor biopsies have been more frequently observed than benign tumor biopsies in both CMT and FMT cases. The resulting data originating from this study can be an aid for veterinary oncologists in better educating clients and planning treatment and prevention strategies and it can be used as a basis for further experimental studies in the oncology section.

Correlation between fine-needle aspiration biopsy and routine histopathology in the diagnosis of spontaneous feline mammary gland tumours

The present study evaluated the correlation between fine-needle aspiration biopsy and routine histopathological examination in the diagnosis of spontaneous mammary gland tumours in cats. From the studied 120 neoplastic growths, cytological diagnosis was correct in 111 cases (92.5%), and incorrect in 9 (7.5%) patients. Out of all studied tumours, 3 false positive (2.5%) and 6 false negative (5%) results were found out. Among benign tumours, cytological diagnosis was correct in 17 cases (94.44 %). Only one false negative diagnosis was demonstrated (5.56% of all benign tumours). As to malignant tumours, cytological diagnosis was correct in 94 (92.15%), and incorrect in 8 patients (7.85%). For diagnostics of feline mammary neoplasms, the sensitivity of cytology was 95.23%, the specificity - 75%, positive predictive value: 97.5% and negative predictive value: 60% compared to routine histopathology. The analysis with Cohen’s kappa test showed agreement of 99.17% (Cohen’s κ 0.96) between cytology and histopathology for benign and 94.17% (Cohen’s κ 0.75) for malignant mammary neoplasms.

Clinicopathological Diversity and Epidemiological Aspects of Canine and Feline Mammary Gland Tumors in Tehran: A Survey (2020-2022)

Background: Mammary tumors are common in dogs and cats. They are models for investigating carcinogenesis and designing treatment protocols that benefit human beings. Senescence, sex, and reproductive status affect the development of such neoplasms. Objectives: In Iran’s absence of a national animal tumor registry, the present study determined clinicopathological and epidemiological aspects of canine and feline mammary tumors in referral cases of four veterinary practices across Tehran from 2020 to 2022. Here, the incidence and types of canine and feline mammary tumors are described, in addition to sex, reproductive status, age, breed, the affected mammary gland(s), grades, lymphatic invasion, and lymph node metastases. Methods: All canine and feline patients with masses in the mammary gland region were considered in this study. The resected tumors and occasional lymph nodes were macroscopically scrutinized. Hematoxylin-Eosin slides were reviewed by light microscopy and immunohistochemistry was utilized when necessary. Results: Of the 76 dogs and eight cats, 100% were females, and the majority were intact. Most patients were 10-11 years old. Purebreds were the most frequent referrals. In some patients, more than one mammary gland had developed neoplasm, i.e. 141 affected glands in 76 bitches and nine affected glands in eight queens. Tumors were presented in both chains and even on the ventral midline, with an increased preponderance of caudally located glands. Thus, the caudal-most glands, i.e. inguinal and caudal abdominal glands, constituted 31.2% and 66.7% of the affected glands in bitches and queens, respectively. Intraductal papillary carcinoma (17%) and complex carcinoma (16.3%) had the highest incidence in dogs. The most commonly observed lesions in cats were lobular hyperplasia with atypia, and duct ectasia, each encompassing 22.3% of the affected glands. Most canine neoplasms were grade I (90.3%), while grade II neoplasms had the highest incidence in cats (50%). Lymphatic invasion and lymph node metastases were seen in canine anaplastic carcinoma, solid carcinoma, and complex carcinoma, as well as feline comedocarcinoma and tubular carcinoma. Conclusion: The risk of developing malignant tumors increases as the individual ages, and generally, caudally located mammary glands in intact purebred females are at increased risk. Moreover, anaplastic carcinomas must be precisely examined, both clinically and histopathologically, for lymph node metastases.

Mammary tumors of the dog and the cat: modern approaches to classification and diagnosis (review)

The study of oncopathology of the mammary gland of cats and dogs is a very relevant problem. This is evidenced by the presence of numerous scientific works and experimental research data of scientists from many countries. For a long time, there were no systematic approaches to the nomenclature and classification of canine and feline mammary gland tumors. That is why different countries of the world had their own national approaches to defining these pathologies. Long-term incomplete analysis of clinical cases, gaps in histopathology and interpretation of the obtained results were reflected in contradictory results, inaccuracies in terminology, definition of diagnosis and interpretations. This also inhibited clinical and experimental research, development and testing of effective drugs. Accumulation of clinical data, consolidated work of morphologists became the basis of lively discussions, which in turn was reflected in decisions to create unified classifications. Histological classification became the basis for the division of tumors of the mammary gland of cats and dogs. When conducting a scientific review, modern data from information resources and information from numerous international symposia and conferences were used. The authors also highlighted the key mechanisms of immune regulation of the oncogenic process. The role of immune cells, mediators of immune protection, in the development of oncopathology of the mammary gland is emphasized. Certain immunological tumor markers have been characterized, which are of great importance in the early stages of diagnosis. Special instrumental methods used by veterinary medicine doctors in routine diagnostics are also described. Prospects for the use of cell therapy with the use of modern immunological technologies were also made. The publication has a review and analytical nature and aims to attract the attention of scientists, diagnosticians and clinicians of various profiles to this issue, which will certainly become a continuation in the study of oncopathology. The unified classification will definitely be useful to scientists and practicing doctors, will enrich their knowledge about the mechanisms of tumor growth. We believe that a unified classification will allow researchers to use informative methods of diagnosis – both as a powerful tool in making an accurate diagnosis, and for effective approaches to differentiation and treatment strategies for patients.

Epithelial-to-Mesenchymal Transition and Phenotypic Marker Evaluation in Human, Canine, and Feline Mammary Gland Tumors

Epithelial-to-mesenchymal transition (EMT) is a process by which epithelial cells acquire mesenchymal properties. EMT has been closely associated with cancer cell aggressiveness. The aim of this study was to evaluate the mRNA and protein expression of EMT-associated markers in mammary tumors of humans (HBC), dogs (CMT), and cats (FMT). Real-time qPCR for SNAIL, TWIST, and ZEB, and immunohistochemistry for E-cadherin, vimentin, CD44, estrogen receptor (ER), progesterone receptor (PR), ERBB2, Ki-67, cytokeratin (CK) 8/18, CK5/6, and CK14 were performed. Overall, SNAIL, TWIST, and ZEB mRNA was lower in tumors than in healthy tissues. Vimentin was higher in triple-negative HBC (TNBC) and FMTs than in ER+ HBC and CMTs (p < 0.001). Membranous E-cadherin was higher in ER+ than in TNBCs (p < 0.001), whereas cytoplasmic E-cadherin was higher in TNBCs when compared with ER+ HBC (p < 0.001). A negative correlation between membranous and cytoplasmic E-cadherin was found in all three species. Ki-67 was higher in FMTs than in CMTs (p < 0.001), whereas CD44 was higher in CMTs than in FMTs (p < 0.001). These results confirmed a potential role of some markers as indicators of EMT, and suggested similarities between ER+ HBC and CMTs, and between TNBC and FMTs.

Histopathology and Immunohistochemistry of tumors in animals attending veterinary teaching hospital

We aim in the current study to investigate the pathology and incidence of tumors that are excised surgically in Veterinary Teaching Hospital. This study collected the tissue samples from animals from 1 October 2020 until 1 April 2021. These samples were collected from animals that undergo surgical procedures to remove neoplastic growths in their body. After tissue grossing, samples were collected from tumor mass, then fixed in neutral buffered 10% formalin for 72 hours, then processed to embedded in paraffin wax. A routine Harris’s hematoxylin and eosin stain were also used, histochemical stains such as Masson’s trichrome and PAS techniques were used as needed. In exceptional cases, IHC protocol was used for diagnostic steps to some types of tumors included in the current study. The antibodies that were used are Vimentin for canine osteosarcoma, P53 for squamous cell carcinoma, Mdm2 for ovine pulmonary adenocarcinoma, and CD10 for neoplastic lymphocytes. The result of the current study showed that the prevalence of tumor recorded was Marek’s diseases 22.8%, squamous cell carcinoma 22.8%, ovine pulmonary adenocarcinoma 15.4%, third eyelid adenoma 15.4%, canine osteosarcomas 7.4%, venereal transmitting tumors 7.4%, and feline mammary gland tumor 7.4%. Histological examination of feline mammary gland tumor composed from server infiltration of neoplastic cells, these neoplastic cells identified as lymphocytes in a fibrous stroma. Third eyelid adenomas in cats and dogs were composed of well-demarcated nodules containing neoplastic sebaceous cells present in glandular form and combined by hyperplasia of epidermis extended inside the neoplastic mass. Venereal transmitting tumor in cat composed from cells with a round to oval shape and the cytoplasm is poorly defined by routine stain, with dense fibrous tissue. Osteosarcoma in a dog has a pleomorphic nature with an anaplastic form composed of islands of osteoid materials, and the tumor is composed of three types of osteoblast, chondroblast, and fibroblast. Pulmonary adenocarcinoma in sheep composed of a nodular formation containing dense collagen fiber deposition with transformation of cells lining alveoli into cuboidal or columnar cells can also be observed in other air passages. Squamous cell carcinomas are primarily found in skin lesions associated with the formation of pathognomic lesions known as keratin pearls. This lesion is composed of the acentric layer of keratin produced by neoplastic keratinocytes. Marek’s disease was observed in visceral organs such as liver, lung, kidney, intestines, and spleen composed from pleomorphic lymphocytes, diffusely proliferating small-to-medium-size lymphoblast. In conclusion, the incidence of tumor in animals that attending Veterinary Teaching Hospital belong to the College of Veterinary Medicine, the University of Mosul was in general high in comparison to other studies in Iraq and other countries, this high incidence should pay attention to the causes of these cancerous conditions and their relation to environmental etiology.

Histopathological and immunohistochemical characteristics of malignant adenomyoepithelioma in a cat: case report

ABSTRACT The malignant adenomyoepithelioma is a rare mammary tumor in women and uncommon in cats with only one report in this species. In this case report, the histopathological and immunohistochemical characteristics of six cases of malignant adenomyopithelioma in the feline mammary gland are described. Microscopic evaluation of tumors showed dense cellular neoplastic proliferation, composed of malignant myoepithelial and epithelial cells, formed by varied arrangements and presenting papillary, tubular and solid nest proliferation. Immunohistochemistry was performed for markers Ki67, Cox-2, RE, RP, p63 and HER-2. All cases were positive for p63, confirming the myoepithelial nature of neoplastic cells. The diagnosis of malignant adenomyopithelioma was made possible through the association between histopathological characteristics and immunohistochemical results.

The evaluation of non-anesthetic computed tomography for detection of pulmonary parenchyma in feline mammary gland carcinoma: a preliminary study

BackgroundThoracic radiography in awake cats is a common procedure for the evaluation of pulmonary metastasis in feline mammary gland carcinoma (MGC). However, due to poor sensitivity, computed tomography (CT) is progressively taking its place. To perform CT in animals, general anesthesia is normally preferred but can cause lung atelectasis, affecting lung interpretation. Besides, MGC is often found in senile cats that are concurrently affected with other diseases, increasing anesthetic risk. Therefore, this study was aimed at comparing the effect of anesthesia on lung atelectasis observed through CT in clinically healthy cats and comparing the feasibility of non-anesthetic CT with non-anesthetic radiography in the detection of lung lesions in feline MGC. Thoracic CTs from anesthetized, clinically healthy cats and non-anesthetized either clinically healthy cats or MGC-affected cats were reviewed. In clinically healthy cats, motion artifacts and characteristics of lung atelectasis were observed and compared. In MGC-affected cats, motion artifacts were observed and compared to clinically healthy cats, and the number of MGC-affected cats, the number and characteristics of lung lesions were compared between non-anesthetic thoracic CT and radiography.ResultsAnesthesia significantly increased lung CT attenuation (P = 0.0047) and was significantly correlated with lung atelectasis (OR = 15; CI 2.02–111.18; P = 0.0081), particularly of the cranial lung lobe. Nonetheless, significantly higher motion artifacts in the caudal thoracic area were found in non-anesthetized healthy cats (P = 0.0146), but comparable low motion artifacts were observed in anesthetized healthy and MGC-affected cats. Non-anesthetic CT revealed higher numbers of MGC-affected cats and pulmonary nodules with a significantly lower nodular diameter (P = 0.0041) than those observed on radiographs. The smallest nodular diameters detected on radiographs and CT were 2.5 and 1.0 mm, respectively. Furthermore, CT showed additional information such as intra-thoracic lymphadenopathy, that could not be seen on radiographs.ConclusionsDespite the motion artifacts, CT without anesthesia is a sensitive technique as it provides better lung inflation. Furthermore, compared to non-anesthetic radiography, non-anesthetic CT provided more information such as higher number of pulmonary nodules of a smaller size, including more distinct intra-thoracic lesions, such as lymphadenopathy, in MGC-affected cats.

Genetic variants of BRCA1 and BRCA2 genes in cats with mammary gland carcinoma.

Mammary tumours are the first and third most incident neoplasm in women and cats, respectively. Approximately 85% of feline mammary gland tumours are malignant and aggressive, especially the triple-negative and HER-2+ molecular subtypes. Triple-negative basal-like feline mammary carcinomas (FMCs) are considered suitable models due to the clinical and morphological similarities with human basal-like triple-negative breast cancer (TNBC). In women, TNBC has a poor prognosis and is often associated with mutations in the tumour suppressor genes BRCA1 and BRCA2. In light of this, the aim of the present investigation was to screen somatic and germline variants of BRCA1 and BRCA2 in nine female cats bearing FMCs. Matched whole blood and FMC samples were obtained for genetic analysis. Additional tumour samples were obtained for histopathological and immunohistochemical evaluation. Genomic DNA was isolated and 27 exonic regions of BRCA1 and BRCA2 genes were amplified and screened by next-generation sequencing. A somatic variant with high functional impact was found in exon 11 of BRCA2 at a frequency of 4.34% in one FMC-bearing cat. Four germline variants with moderate impact were detected in three of the nine FMC-bearing cats and were restricted to exon 9 of BRCA1. It is concluded that the germline genetic variants found in one-third of FMC-bearing animals might be associated with a higher risk of hereditary mammary carcinogenesis.

The anti-proliferative and apoptotic effects of curcumin on feline mammary gland tumor cells in vitro.

Feline mammary gland tumors (FMGTs) are the third most diagnosed tumors in cats. Feline mammary gland tumors have aggressive biological behavior and poor response to both surgical and medical treatments, thus, new therapeutic approaches are essential to improve. Curcumin (CUR) is a polyphenol component exhibiting anti-cancer effects and induces apoptosis through different mechanisms especially in human breast cancer. However, there is no study investigating the effects of CUR on FMGTs. The aim of this study was to determine the anti-proliferative and apoptotic effects of CUR on primary cell lines from FMGT tissue samples of two cases classified as carcinoma-simple, tubular type (grade III). The cytotoxic effect of CUR was determined by water-soluble tetrazolium salt-1 (WST-1) assay. Annexin V, cell cycle, and acridine orange (AO) analyses were performed to determine the apoptotic effect of CUR. Our results showed that CUR had an anti-proliferative and apoptotic effect through induction of apoptosis and cell cycle arrest (G0/G1) on FMGT cells. Therefore, this is the first study that shows the effects of CUR on FMGTs. However, further molecular studies are required to compare the effects of CUR on different histopathological phenotypes and to determine the further molecular mechanisms including the potential apoptotic and cellular pathways affected by CUR.

Expression of HIF-1\u03b1 and VEGF in feline mammary gland carcinomas: association with pathological characteristics and clinical outcomes

BackgroundThe microenvironment within solid malignant tumors, including feline mammary gland carcinomas (FMGCs), is commonly hypoxic, possibly due to the lack of functional blood vessels in rapidly proliferating neoplastic tissue. Malignant cells can undergo genetic and adaptive changes that prevent them from dying due to oxygen deprivation through expressions of hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF). Therefore, HIF-1α and VEGF are ideal biomarkers for cancer therapy and prognostic evaluation. The aims of this study were to evaluate the expression of HIF-1α and VEGF in feline mammary carcinomas and analyze their correlations with clinical and pathological factors, such as clinical stage, histologic grading, regional metastasis, and overall survival rate.ResultsParaffin-embedded tissue samples collected from 72 cats with FMGCs were retrospectively studied. Histologic pattern and histologic grading (Elston and Ellis grading system) of these FMGCs were determined. Our data indicated that grade II tubulopapillary carcinomas (43/72, 59.7%) prevailed in this study, and most FMCGs showed apparent necrosis, squamous metaplasia, and intratumoral stromal response. According to the results of immunohistochemical (IHC) stainings performed in tissue microarrays (TMAs), HIF-1α and VEGF overexpressions were respectively noted in 69.4% (50/72) and 77.8% (56/72) of FMGC cases. Chi-square test showed no correlation of HIF-1α overexpression with clinical and pathological factors. VEGF overexpression was significantly correlated with histologic pattern (p = 0.021), stromal response (p = 0.048), squamous metaplasia (p = 0.001), and lymphovascular invasion (p = 0.007). However, neither HIF-1α nor VEGF overexpression was correlated with histologic grading and metastasis. Of 38 cats with 1-year follow-up, IHC stainings of HIF-1α and VEGF were performed on whole tissue sections. The results showed that overexpression of HIF-1α was significantly correlated with the overall survival rate (p < 0.05) (log-rank test), whereas there was no significant correlation between VEGF overexpression and overall survival rate.ConclusionsThis study suggests that the overexpression of HIF-1α may indicate poor prognosis/overall survival rate in cats with FMGCs. Developing compounds that inhibit HIF-1α may be a potential approach to FMGC treatment.

Histological Characterization of Some Feline Mammary Gland Tumors with Whole Slide Images Scan as a Trial of Remote Diagnosis

Histological Characterization of Some Feline Mammary Gland Tumors with Whole Slide Images Scan as a Trial of Remote Diagnosis

Mouse mammary tumour virus-like env nucleotide and p14 signal peptide are present in feline mammary carcinomas, but not in neoplastic or dysplastic canine mammary lesions.

Mouse mammary tumour virus-like (MMTV-like) is suspected to be involved in human breast cancer and it has been hypothesized that companion animals might have a role in viral transmission. The aim of our study was to investigate the presence of MMTV-like nucleotide sequences and viral protein in a larger number of feline (FMCs) and canine mammary carcinomas (CMCs) by nested PCR and immunohistochemistry. Results showed that the presence of MMTV-like env sequence in FMCs was 7% (6/86), while all the CMCs and canine dysplastic lesions scored negative. All PCR-positive FMCs scored positive for the MMTV p14 signal peptide of the envelope precursor protein of the virus. In contrast, all PCR-negative FMCs and canine mammary lesions were also negative for immunohistochemistry analysis. Canine and feline normal mammary gland tissues scored negative for both PCR and MMTV-p14 protein. Multiple nucleotide alignment of MMTV-like env gene sequences isolated from cat showed 97% and 99% similarity with HMTV and MMTV, respectively, while the others two presented some polimorphisms. Particularly the sequences of one of these two tumors showed a polymorphism (c.7575 A> G), that causes a previously unreported amino acid substitution (Thr > Ala). In conclusion, the results of our study showed the presence of MMTV-like sequences and viral protein in some FMCs. Further studies are needed to understand whether this virus does play a role in the development of FMCs, if MMTV-like is an exogenous virus as these data suggest and, in such a case, how and from whom this virus was acquired.

Expression of iron-related proteins in feline and canine mammary gland reveals unexpected accumulation of iron

Dysregulation of cellular iron homeostasis in human breast cancer is reflected by the altered expression of regulatory proteins. The expressions of iron-related proteins in the mammary glands of cats and dogs have not been assessed. We evaluated the expressions of ferritin, ferroportin, hepcidin and transferrin receptor 1 in benign and malignant mammary gland lesions in cats and dogs. Iron deposition was detected using Perls’ Prussian blue staining. We found no major differences in the expression of iron-related proteins between benign and malignant mammary gland lesions in either cats or dogs; however, these species exhibited accumulation of iron in benign lesions. Our findings provide an explanation for the absence of higher iron requirements by tumor cells in these animals. Further investigation of local iron homeostasis in cats and dogs and differences in their physiology compared to human breast cancer is required.

Evaluation of protein kinase CK2 as a therapeutic target for squamous cell carcinoma of cats.

OBJECTIVE To investigate protein kinase CK2 (CK2) expression in squamous cell carcinoma (SCC) of cats and to examine effects of CK2 downregulation on in vitro apoptosis and viability in SCC. SAMPLE Biopsy specimens of oral mucosa and testis and blood samples from clinically normal cats, biopsy specimens of oral SCC from cats, and feline SCC (SCCF1) and mammary gland carcinoma (K12) cell lines. PROCEDURES Immunohistochemical labeling for CK2α was performed on biopsy specimens. Sequences of the CK2α subunit gene and CK2α' subunit gene in feline blood and feline cancer cell lines were determined by use of PCR and reverse-transcription PCR assays followed by direct Sanger sequencing. Specific small interfering RNAs (siRNAs) were developed for feline CK2α and CK2α'. The SCCF1 cells were treated with siRNA and assessed 72 hours later for CK2α and CK2α' expression and markers of apoptosis (via western blot analysis) and for viability (via 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium assays). RESULTS CK2α was expressed in all feline oral mucosa samples and 7 of 8 oral SCC samples. Expression of CK2α and CK2α' was successfully downregulated in SCCF1 cells by use of siRNAs, which resulted in decreased viability and induction of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE In this study, CK2 appeared to be a promising therapeutic target for SCCs of cats. A possible treatment strategy for SCCs of cats would be RNA interference that targets CK2.

Retraction Note: Comparative value of clinical, cytological, and histopathological features in feline mammary gland tumors; an experimental model for the study of human breast cancer.

The diagnosis of breast lesions is usually confirmed by fine-needle aspiration cytology (FNAC) or histological biopsy. Although there is increasing literature regarding the advantages and limitations of both modalities, there is no literature regarding the accuracy of these modalities for diagnosing breast lesions in high-risk patients, who usually have lesions detected by screening. Moreover, few studies have been published regarding the cytopathology of mammary tumors in cats despite widespread use of the animal model for breast cancer formation and inhibition. The objective of the present study was to evaluate the diagnostic interest of cytological and histopathological analysis in feline mammary tumours (FMTs), in order to evaluate its possible value as an animal model. The study was performed in 3 female cats submitted to surgical resections of mammary tumours. The mammary tumours were excised by simple mastectomy or regional mastectomy, with or without the superficial inguinal lymph nodes. Female cats were of different breeds (1 siamese and 2 persians). Before surgical excision of the tumour, FNA cytology was performed using a 0.4 mm diameter needle attached to a 8 ml syringe held in a standard metal syringe holder. The cytological sample was smeared onto a glass slide and either air-dried for May-Grunwald-stain and masses were surgically removed, the tumours were grossly examined and tissue samples were fixed in 10%-buffered-formalin and embedded in paraffin. Sections 4 μm thick were obtained from each sample and HE grade III, (2/3) with high mitotic index. The preferential localization of mammary neoplasms was in the inguinal lobe (1/3 case) and abdominal lobes (2/3 cases). Furthermore, 1case of the inguinal mass affected the left caudo-inguinal lobe and 2cases right cranio and caudo abdominal lobes. The study concluded that cytology could be used as a quick, rapid, field diagnostic technique in combination with histopathology for the diagnosis of feline mammary tumors (FMTs). Our findings in feline MTs indicate that FMTs could be useful as an animal model of human breast cancer. Moreover, because of the similarity of the cytohistopathological findings in the human and feline mammary gland tumours, it is possible to use the same cytopathological criteria applied in human pathology for the diagnosis of feline mammary gland tumours. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2047361423103295

Properties of the feline tumour suppressor reduced expression in immortalized cells (REIC/Dkk-3).

Reduced expression in immortalized cells (REIC/Dkk-3), a member of the human Dickkopf (Dkk) family, is a growth suppressor in human and canine mammary tumours. Mammary gland tumours are common neoplasms with high malignancy in female cats. The purpose of this study was to clone the feline REIC/Dkk-3 homolog, investigate its expression in cell lines established from feline mammary gland tumours, and test its tumour suppressor function. Western blot analysis revealed that expression of the REIC/Dkk-3 protein was reduced in feline mammary carcinoma cell lines. Forced expression of REIC/Dkk-3 induced apoptosis in feline mammary tumour cell lines. These results demonstrate that REIC/Dkk-3 expression, which is downregulated in feline mammary tumour cell lines, results in the induction of apoptosis in these cells. Our findings suggest that feline REIC/Dkk-3 represents a potential molecular target for the development of therapies against feline mammary cancers.

An Immunohistochemical Study of Epithelial–Mesenchymal Transition in Feline Mammary Tumours

Epithelial-mesenchymal transition (EMT), as defined by loss of epithelial characteristics and gain of a mesenchymal phenotype, has been reported in human and canine mammary tumors, but similar study in feline mammary tumors has not been performed. EMT can be induced or regulated by various growth and differentiation factors among which TGFβ has received much attention. The aim of our study was to evaluate loss of epithelial and gain of mesenchymal markers in epithelial neoplastic cells of feline mammary tumors and to assess possible correlation with expression of TGFβ. Materials and Methods: A total of 27 feline mammary gland hyperplastic and neoplastic lesions were selected from archive of Department of veterinary pathology Zagreb (4 benign lesions, 3 grade I carcinomas, 8 grade II and 12 grade III carcinomas) to evaluate immunohistochemical expression of cytokeratin AE1/AE3, E-cadherin, vimentin and TGFβ. One-way ANOVA with post hoc Tukey's test and Spearman's rank correlation coefficient were used for statistical analysis. Results: Generally, loss of cytokeratin and E-cadherin, and gain of vimentin were more frequently observed in carcinomas of higher malignancy grade compared to benign lesions. However, the correlation between each marker showed only positive correlation between loss of E-cadherin and gain of vimentin (r=0.69, p<0.05) in grade II carcinomas. The expression of TGFβ was significantly higher in carcinomas grade II and III compared to grade I (p<0.05). Conclusions: These findings indicate occurrence of characteristic changes suggestive of EMT in feline mammary carcinomas and a possible role of TGFβ in this process.

Caracterização clínica e terapêutica de lesões mamárias não neoplásicas em fêmeas da espécie felina

The fibroepithelial hyperplasia and cystic dysplasia correspond to feline mammary gland nonneoplastic proliferations. Although benign, they have the carcinoma as the main differential diagnosis. The study aimed to establish the clinical-epidemiological and therapeutic profile of nonneoplastic mammary neoformations of domestic cats, in order to help veterinary clinicians in the management of feline mastopathies. Information was obtained retrospectively from the Veterinary Hospital of the Semi-Arid Rural Federal University clinical records. Historical information, clinical appearance of the lesions and microscopic diagnosis were collected, besides the therapy used and the respective clinical response. The data were distributed in frequencies. It was found that most disorders (91%) corresponded to fibroepithelial hyperplasia, in contrast with one (9%) case of cystic dysplasia. Cytology was enough to confirm all cases of fibroepithelial hyperplasia, while the cystic dysplasia definitive diagnosis was obtained only by the histopathological exam. Many of the cats studied were under one year old, having not usually been ovariectomized yet, but already subjected to progestin treatment. The most common clinical sign were local cyanosis. In 90% of the female patients with breast fibroepithelial hyperplasia, primary treatment consisted of antiprogestagen drug (aglepristone) administration. After complete regression of the breast tissue, the patients underwent ovariosalpingohysterectomy. For the cystic dysplasia case, it was established the association of surgical sterilization with mastectomy. For all situations, the different therapies promoted permanent remission of the mastopathies. The data obtained become crucial for clinical management and therapy of feline patients suffering from nonneoplastic mammary gland neoformations.