Establishment and Characterization of Canine Mammary Gland Carcinoma Cell Lines With Vasculogenic Mimicry Ability in vitro and in vivo

Lainetti, Patrícia de Faria; Brandi, Andressa; Leis-Filho, Antonio Fernando; Prado, Maria Carolina Mangini; Kobayashi, Priscila Emiko; Laufer-Amorim, Renée; Fonseca‐Alves, Carlos Eduardo

doi:10.3389/fvets.2020.583874

Cited by 14 publications

(16 citation statements)

References 66 publications

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“…After achieving 80% confluence in the bottle, the cells were trypsinized and placed in new 75 cm² bottles with a filter. The cell lines used, UNESP-CM1 and UNESP-CM9, were previously established and characterized by Lainetti [ 36 ]. UNESP-CM1 comes from a 12-year-old poodle and was classified as a solid carcinoma, grade II with tubular formation and HER2 overexpressing.…”

Section: Methodsmentioning

confidence: 99%

Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines

Pedro,

Brasileiro,

Macedo

et al. 2024

J. Venom. Anim. Toxins incl. Trop. Dis

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Background: Mammary gland tumors are the most prevalent neoplasm in intact female dogs, and they are good natural models to study comparative oncology. Most canine mammary malignancies, as in women, are commonly refractory to conventional therapies and demand continuous new therapeutic approaches. Crotalus durissus terrificus , also called rattlesnake, has more than 60 different proteins in its venom with multiple pharmaceutical uses, such as antitumor, antiviral, and antimicrobial action. Crotoxin, a potent β-neurotoxin formed by the junction of two subunits, a basic subunit (CB-PLA 2 ) and an acidic subunit (crotapotin), has already been reported to have anticancer properties in different types of cancers. Methods: In this work, we describe the cytotoxic potential of crotoxin and its subunits compared to doxorubicin (drug of choice) in two canine mammary carcinoma cell lines. Results: Crotoxin, CB-PLA 2 , crotalic venom, and doxorubicin decreased cell viability and the ability to migrate in a dose-dependent manner, and crotapotin did not present an antitumoral effect. For all compounds, the predominant cell death mechanism was apoptosis. In addition, crotoxin did not show toxicity in normal canine mammary gland cells. Conclusion: Therefore, this work showed that crotoxin and CB-PLA 2 had cytotoxic activity, migration inhibition, and pro-apoptotic potential in canine mammary gland carcinoma cell lines, making their possible use in cancer research.

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Section: Methodsmentioning

confidence: 99%

Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines

Pedro,

Brasileiro,

Macedo

et al. 2024

J. Venom. Anim. Toxins incl. Trop. Dis

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“…Four tumors previously characterized [ 71 ] were used to perform immunofluorescence for AKT, PTEN, mTOR and 4EBP1 antibodies. Then, tumor cells were expanded and analyzed for cell viability (MTT), gene expression (RT-qPCR) and proteomics as can be observed in Figure 6 .…”

Section: Methodsmentioning

confidence: 99%

“…In immunohistochemistry, primary tumors were both classified as HER-2 overexpressing. Grade II solid carcinoma bone metastasis was classified as triple negative with non-basal immunophenotype and adenosquamous carcinoma lymph node metastasis as HER-2 overexpressing, previously described by our group in Lainetti et al [ 71 ].…”

Section: Methodsmentioning

confidence: 99%

Proteomics Approach of Rapamycin Anti-Tumoral Effect on Primary and Metastatic Canine Mammary Tumor Cells In Vitro

et al. 2021

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Rapamycin is an antifungal drug with antitumor activity and acts inhibiting the mTOR complex. Due to drug antitumor potential, the aim of this study was to evaluate its effect on a preclinical model of primary mammary gland tumors and their metastases from female dogs. Four cell lines from our cell bank, two from primary canine mammary tumors (UNESP-CM1, UNESP-CM60) and two metastases (UNESP-MM1, and UNESP-MM4) were cultured in vitro and investigated for rapamycin IC50. Then, cell lines were treated with rapamycin IC50 dose and mRNA and protein were extracted in treated and non-treated cells to perform AKT, mTOR, PTEN and 4EBP1 gene expression and global proteomics by mass spectrometry. MTT assay demonstrated rapamycin IC50 dose for all different tumor cells between 2 and 10 μM. RT-qPCR from cultured cells, control versus treated group and primary tumor cells versus metastatic tumor cells, did not shown statistical differences. In proteomics were found 273 proteins in all groups, and after data normalization 49 and 92 proteins were used for statistical analysis for comparisons between control versus rapamycin treatment groups, and metastasis versus primary tumor versus metastasis rapamycin versus primary tumor rapamycin, respectively. Considering the two statistical analysis, four proteins, phosphoglycerate mutase, malate dehydrogenase, l-lactate dehydrogenase and nucleolin were found in decreased abundance in the rapamycin group and they are related with cellular metabolic processes and enhanced tumor malignant behavior. Two proteins, dihydrolipoamide dehydrogenase and superoxide dismutase, also related with metabolic processes, were found in higher abundance in rapamycin group and are associated with apoptosis. The results suggested that rapamycin was able to inhibit cell growth of mammary gland tumor and metastatic tumors cells in vitro, however, concentrations needed to reach the IC50 were higher when compared to other studies.

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“…The expression of cytokeratins and vimentin in breast cancer is linked to malignant tumor (Heatley et al, 1993). Although markers were employed and referred to prior investigations, this work has not yet completed on additional markers that can be used for confirmation (de Faria Lainetti et al, 2020). For example, vasculogenic mimicry (VM) proteins include VEcadherin, tumor groeth factor 1 (TGF-1), and EpCam.…”

Section: Case 3: Histiocytic Positive Expression Of Vimentin In Breas...mentioning

confidence: 99%

Characterisation and Specific Markers Expression of Soft Tissue Sarcoma: Case Reports for the Dog at the Ho Chi Minh City Veterinary Clinic

Nguyen¹,

Thi²,

Linh³

et al. 2022

J. tech. innov.

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Treatment failure in removing canine soft tissue tumors, a locally invasive mesenchymal neoplasm, may occur in between 7 and 75 percent of patients, with recurrence consistently associated with lower overall survival for the dog. This study was to classify the tumor examine the occurrence of cancers in dogs reported at the Ho Chi Minh Veterinary Clinic by integrating blood physiological and histopathological of patient samples. In this study, the percentage of tissue sarcoma in the dog was 1.5%. Histopathological analysis characteristics of the ear lobe tumors displayed a strong proliferation of rhabdoid cells, presence of leukocytes infiltrating around blood vessels, and vaginal tumors showed normal tumor cell density with a strong collagen fibers formation in fibroblast cells and no strong proliferation and ferocity of cells. In particular, immuno-histopathological analysis of biopsies sample from breast tumor showed an overexpressing of vimentin marker (68.1%) in cells which indicated status of malignant metastasis. The metastatic tumor cells in breast tumor might cause the change in blood biochemical parameters together with sepsis, thus the dog was not eligible for surgery. In general, histological examination and expression levels of markers can help determine the malignant status of tumors in dogs and should be included medical examination to limit local recurrence following surgical resection.

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Establishment and Characterization of Canine Mammary Gland Carcinoma Cell Lines With Vasculogenic Mimicry Ability in vitro and in vivo

Cited by 14 publications

References 66 publications

Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines

Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines

Proteomics Approach of Rapamycin Anti-Tumoral Effect on Primary and Metastatic Canine Mammary Tumor Cells In Vitro

Characterisation and Specific Markers Expression of Soft Tissue Sarcoma: Case Reports for the Dog at the Ho Chi Minh City Veterinary Clinic

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