Preliminary investigation of extracellular vesicles in mammary cancer of dogs and cats: Identification and characterization

Sammarco, Alessandro; Finesso, G.; Cavicchioli, Laura; Ferro, Silvia; Caicci, Federico; Zanetti, Rossella; Sacchetto, Roberta; Zappulli, Valentina

doi:10.1111/vco.12405

Cited by 17 publications

(12 citation statements)

References 51 publications

(140 reference statements)

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“…In veterinary medicine, very little is known on cancer‐derived EVs. There is only a preliminary investigation on extracellular vesicles in canine and feline mammary cancer [72]. Further studies are necessary to isolate, identify and characterize EVs from IBC/IMC cell lines.…”

Section: Discussionmentioning

confidence: 99%

Vasculogenic mimicry-associated ultrastructural findings in human and canine inflammatory breast cancer cell lines

et al. 2019

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Background Human inflammatory breast cancer (IBC) and canine inflammatory mammary cancer (IMC) are the most lethal mammary cancers. An exacerbated angiogenesis and the existence of vasculogenic mimicry (VM) are hallmarks of these tumors. The information regarding VM and ultrastructural characteristics of mammary cell lines is scant. Methods In this study, IBC cell line SUM149 and IMC cell line IPC-366 in adherent (2D) and non-adherent (3D) (mammospheres, cancer stem cells) conditions were analyzed by transmission and scanning electron microscopy (TEM and SEM, respectively). Results The TEM revealed round to oval shape cells with microvilli on the surface, high numbers of peroxisomes in close apposition to lipid droplets and some extracellular derived vesicles. The TEM and the SEM mammospheres revealed group of cells clumping together with a central lumen (resembling a mammary acini). The cells joint are tight junctions and zonula adherens. By SEM two cell morphologies were observed: spherical and flattened cells. There was evidence endothelial-like cells (ELCs), which is characteristic for this disease, showing several or unique cytoplasmic empty space. ELCs were more frequent in 3D than in 2D culture conditions and contained Weibel-Palade cytoplasmic bodies, which are exclusive structures of endothelial cells. Conclusions Both cell lines, IPC-366 and SUM-149, shared ultrastructural characteristics, further supporting canine IMC as a model for the human disease. To the best of our knowledge, this is the first study that demonstrate the morphological differentiation of cultured cancer stem cells from cancer epithelial cell lines into endothelial-like cells, confirming the vasculogenic mimicry phenomenon from an ultrastructural point of view.

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Section: Discussionmentioning

confidence: 99%

Vasculogenic mimicry-associated ultrastructural findings in human and canine inflammatory breast cancer cell lines

et al. 2019

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“…The same study also used transmission immuno-electron microscopy using immunogold-labeled antibodies to common EVs markers (CD63 and Alix). 171 A limitation of this technique is the limited number of EVs that can be analyzed. For this reason, it is impossible to use electron microscopy to analyze a representative population of EVs that vary in size and composition, as is present in biological and clinical samples.…”

Section: Methods For Isolating and Detecting Evsmentioning

confidence: 99%

Extracellular Vesicles: Novel Opportunities to Understand and Detect Neoplastic Diseases

et al. 2021

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With a size range from 30 to 1000 nm, extracellular vesicles (EVs) are one of the smallest cell components able to transport biologically active molecules. They mediate intercellular communications and play a fundamental role in the maintenance of tissue homeostasis and pathogenesis in several types of diseases. In particular, EVs actively contribute to cancer initiation and progression, and there is emerging understanding of their role in creation of the metastatic niche. This fact underlies the recent exponential growth in EV research, which has improved our understanding of their specific roles in disease and their potential applications in diagnosis and therapy. EVs and their biomolecular cargo reflect the state of the diseased donor cells, and can be detected in body fluids and exploited as biomarkers in cancer and other diseases. Relatively few studies have been published on EVs in the veterinary field. This review provides an overview of the features and biology of EVs as well as recent developments in EV research including techniques for isolation and analysis, and will address the way in which the EVs released by diseased tissues can be studied and exploited in the field of veterinary pathology. Uniquely, this review emphasizes the important contribution that pathologists can make to the field of EV research: pathologists can help EV scientists in studying and confirming the role of EVs and their molecular cargo in diseased tissues and as biomarkers in liquid biopsies.

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“…Cell protein concentration was calculated using Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). In order to evaluate CCND1 antibody specificity, a western blot analysis has been performed as previously described . Twenty‐five micrograms of proteins from cells were denatured at 70°C for 10 minutes.…”

Section: Methodsmentioning

confidence: 99%

Cyclin D1 immunohistochemical expression and somatic mutations in canine oral melanoma

Zamboni

Brocca

Ferraresso

et al. 2019

Vet Comparative Oncology

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Canine oral melanoma (COM) is the most frequent tumour with oral localization in dogs. Copy number gains and amplifications of CCND1, a gene coding for Cyclin D1, are the most frequent chromosomal aberrations described in human non-UV induced melanomas. Twenty-eight cases of COM were retrieved from paraffin-blocks archives. A total of 4 μm thick sections were immunostained with an antibody against human Cyclin D1 and Ki-67. Cyclin D1 and Ki-67 expressions were scored through two counting methods. DNA was extracted from 20 μm thick sections of formalin-fixed paraffin-embedded blocks. Pathological and surrounding healthy tissue was extracted independently. Cyclin D1 immunolabelling was detected in 69% (18/26) while Ki-67 was present in 88.5% (23/26) of cases. Statistical analysis revealed correlation between two counting methods for Cyclin D1 (r = 0.54; P = .004) and Ki-67 (r = 0.56; P = .003). The correlation found between Ki-67 and Cyclin D1 indexes in 16/26 cases labelled by both antibodies (r = 0.7947; P = .0002) suggests a possible use of Cyclin D1 index as prognostic marker. Polymerase chain reaction analysis on CCND1 coding sequence revealed the presence of nine somatic mutations in seven samples producing synonymous, missense and stop codons. Since none of the single-nucleotide polymorphisms was found to be recurrent, it is suggested that overexpression of Cyclin D1 may be the consequence of alterations of CCND1 upstream regions or other genetic aberrations not detectable with the methodology used in this study. Future studies are needed to verify the potential use of Cyclin D1 index as prognostic indicator and to highlight the molecular events responsible for Cyclin D1 overexpression in COMs. K E Y W O R D S CCND1, Cyclin D1, dog, immunohistochemistry, melanoma

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Preliminary investigation of extracellular vesicles in mammary cancer of dogs and cats: Identification and characterization

Cited by 17 publications

References 51 publications

Vasculogenic mimicry-associated ultrastructural findings in human and canine inflammatory breast cancer cell lines

Vasculogenic mimicry-associated ultrastructural findings in human and canine inflammatory breast cancer cell lines

Extracellular Vesicles: Novel Opportunities to Understand and Detect Neoplastic Diseases

Cyclin D1 immunohistochemical expression and somatic mutations in canine oral melanoma

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