PDGFR‐α, PDGFR‐β, VEGFR‐2 and CD117 expression in canine mammary tumours and evaluation of the in vitro effects of toceranib phosphate in neoplastic mammary cell lines

Gattino, Francesca; Maniscalco, Lorella; Iussich, Selina; Biasato, Ilaria; Martano, Marina; Morello, Emanuela; Gola, Cecilia; Ruiz, Yolanda Millán; Saeki, Nobuo; Buracco, Paolo; Mulas, J. Martı́n de las; Maria, Raffaella De

doi:10.1136/vr.104414

Cited by 7 publications

(9 citation statements)

References 49 publications

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“…All of them showed a heterogeneous amount of mRNA, indicating cell‐line specific gene expression. Interestingly, similar variations were previously described when considering protein levels …”

Section: Discussionsupporting

confidence: 83%

“…Interestingly, similar variations were previously described when considering protein levels. 6,7,33 Cell lines were further investigated with several biological assays to determine whether cell migration was affected by treatment with TOC. Transwell assays demonstrated that all OSA cell lines were able to migrate in the absence of chemo-attractive factors; conversely, Overall, these in vitro results suggested that Wall and Penny cells responded to TOC treatment mainly by reducing invasion rather than proliferation.…”

Section: Discussionmentioning

confidence: 99%

“…Immunohistochemical evaluation of caspase 3 was carried out by taking images with a 40× microscope objective from 10 random fields per tumour to provide an average index of caspase 3‐positive cells . For the IHC evaluation of c‐Kit, PDGFR‐α, PDGFR‐β and VEGFR‐2 expression, the percentage of positively stained tumour cells and the relative intensity were scored according to the literature …”

Section: Methodsmentioning

confidence: 99%

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In vitro and in vivo effects of toceranib phosphate on canine osteosarcoma cell lines and xenograft orthotopic models

Sánchez-Céspedes

Accornero

Miretti

et al. 2019

Vet Comparative Oncology

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Canine osteosarcoma (OSA) is the most common primary malignant bone tumour in dogs, and it has a high metastatic rate and poor prognosis. Toceranib phosphate (TOC; Palladia, Zoetis) is a veterinary tyrosine kinase inhibitor that selectively inhibits VEGFR‐2, PDGFRs and c‐Kit, but its efficacy is not yet fully understood in the treatment of canine OSA. Here, we evaluated the functional effects of TOC on six OSA cell lines by transwell, wound healing and colony formation assays. Subsequently, two cell lines (Wall and Penny) were selected and were inoculated in mice by intrafemoral injection to develop an orthotopic xenograft model of canine OSA. For each cell line, 30 mice were xenografted; half of them were used as controls, and the other half were treated with TOC at 40 mg/kg body weight for 20 days. TOC inhibited cell growth of all cell lines, but reduced invasion and migration was only observed in Penny and Wall cell lines. In mice engrafted with Penny cells and subjected to TOC treatment, decreased tumour growth was observed, and PDGFRs and c‐Kit mRNA were downregulated. Immunohistochemical analyses demonstrated a significant reduction of Ki67 staining in treated mice when compared to controls. The results obtained here demonstrate that TOC is able to slightly inhibit cell growth in vitro, while its effect is evident only in a Penny cell xenograft model, in which TOC significantly reduced tumour size and the Ki67 index without modifying apoptosis markers.

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Section: Discussionsupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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In vitro and in vivo effects of toceranib phosphate on canine osteosarcoma cell lines and xenograft orthotopic models

Sánchez-Céspedes

Accornero

Miretti

et al. 2019

Vet Comparative Oncology

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“…Normal and hyperplastic mammary tissues show a moderate to strong diffusely cytoplasmic and membranous c-Kit positivity even though the relationship between the tumour histotype and the presence or absence of -c-Kit expression is still debated. Recently published studies have demonstrated a decrease in c-KIT expression in benign tumours as well as in the tubulopapillary carcinoma; no c-KIT expression is reported in high-grade carcinoma, although a cytoplasmic expression pattern could partially increase cellular proliferation (Brunetti et al, 2014;Gattino et al, 2018;Morini et al, 2004). This case report aims to add an extra contribution to the Veterinary Medicine literature on collision tumours.…”

Section: Discussionmentioning

confidence: 82%

Mast Cell Tumour and Mammary Gland Carcinoma Collision Tumour. Case report and literature review.

Rifici¹,

Sfacteria²,

Giorgio³

et al. 2022

J Hellenic Vet Med Soc

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Collision tumours are the coexistence, at the same venue, of distinct tumours not macroscopically distinguishable and consisting of two independent cell populations without histological admixture. In human medicine, collision tumours in different anatomical sites have been described. In the veterinary literature, few cases exist so far. A 12-year-old female Labrador with a mammary gland nodular lesion was presented for clinical examination. The nodule was surgically removed and underwent histological and immunohistochemical analysis. Histopathological examination revealed two distinct malignant tumours: a mammary gland carcinoma and a cutaneous mast cells tumour. To the author's knowledge, the paper reports the first case of a collision tumour composed of mammary gland neoplasia and mast cell tumour. The rising interest in collision tumours suggests widening their knowledge and setting up a multimodal approach that includes surgery and targeted therapy.

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“…The rabbit polyclonal anti‐VEGFR‐2 antibody was previously confirmed to cross‐react with the canine tissue samples by Western blotting and immunohistochemistry 16 and the information about the cluster of differentiation 31 (CD31) antibody cross‐reactivity was provided by the manufacturer (Thermo Fisher Scientific). In this study, the VEGF‐A cross‐reactivity in dogs was evaluated by Western blotting, following the procedures described by Rivera‐Calderón et al 17 Briefly, four canine normal frozen tissues were mechanically homogenized and protein extraction was performed using the radioimmunoprecipitation assay buffer.…”

Section: Methodsmentioning

confidence: 99%

Expression and prognostic significance of vascular endothelial growth factor‐A (VEGF‐A) and its receptor in canine prostate cancer

et al. 2021

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Background Vascular endothelial growth factor‐A (VEGF‐A) and its receptor, VEGF receptor‐2 (VEGFR‐2), represent a complex family of angiogenic molecules consisting of different ligands and receptors. Due to the importance of VEGF‐A/VEGFR‐2 signaling in tumor proliferation and angiogenesis, this study aimed to evaluate the protein and gene expression levels of VEGF‐A and VEGFR‐2 in canine prostate cancer (PC). Methods We analyzed VEGF‐A and VEGFR‐2 expression in 87 PC samples by immunohistochemistry and quantitative‐polymerase chain reaction. PC samples were graded according to the Gleason score and the immunohistochemical staining for VEGF‐A and VEGFR‐2 was quantified using a selected threshold from the ImageJ Software. Microvascular density was assessed by cluster of differentiation 31 staining and counting the number of positive vessels. Additionally, the homology of VEGF‐A and VEGFR‐2 between humans and dogs was assessed, followed by the construction of a protein structure homology model to compare the tertiary structures of these proteins in both species. Results Negative to weakly positive expression levels of VEGF‐A and VEGFR‐2 were observed in the epithelial cells of the normal prostate (NP) and prostatic hyperplasia samples. In contrast, the canine proliferative atrophy and PC samples exhibited higher VEGF‐A (p < .0001) and VEGFR‐2 (p < .0001) compared to NP. Moreover, positive correlations between the expression levels of VEGF‐A and VEGFR‐2 (Spearman's coefficient (r) = .68, p = .013) and the expression levels of VEGF‐A and VEGFR‐2 proteins (r = .8, p < .0001) were also observed in the NP samples. Additionally, the patients with PC exhibiting higher VEGFR‐2 expression levels experienced a shorter survival period (p = .0372). Furthermore, we found an association between the microvascular density and overall survival. Dogs with a higher number of vessels showed a shorter survival time. We further demonstrated that the VEGF‐A and VEGFR‐2 exhibited high homology between humans and dogs, and identified their protein structures in both species. Conclusions In conclusion, VEGFR‐2 appears to be an independent prognostic factor in animals with PC. VEGF‐A and VEGFR‐2 are highly conserved between humans and dogs, which can be investigated further in future cross‐species studies to explore their therapeutic applications.

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PDGFR‐α, PDGFR‐β, VEGFR‐2 and CD117 expression in canine mammary tumours and evaluation of the in vitro effects of toceranib phosphate in neoplastic mammary cell lines

Cited by 7 publications

References 49 publications

In vitro and in vivo effects of toceranib phosphate on canine osteosarcoma cell lines and xenograft orthotopic models

In vitro and in vivo effects of toceranib phosphate on canine osteosarcoma cell lines and xenograft orthotopic models

Mast Cell Tumour and Mammary Gland Carcinoma Collision Tumour. Case report and literature review.

Expression and prognostic significance of vascular endothelial growth factor‐A (VEGF‐A) and its receptor in canine prostate cancer

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