Rapid multiple immunofluorescent staining for the simultaneous detection of cytokeratin and vimentin in the cytology of canine tumors

Sawa, Mariko; Yabuki, Akira; Kohyama, Moeko; Miyoshi, Noriaki; Yamato, Osamu

doi:10.1111/vcp.12598

Cited by 8 publications

(12 citation statements)

References 12 publications

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“…This finding is in total agreement with an early report on the consistent detection of Vimentin expression in six cell lines explanted from COMM (including Shadow), a skin melanoma‐derived cell line and 28/29 tumour sections prepared from 11 COMM, 14 cutaneous and 4 digital canine melanomas . Vimentin is generally expressed by mesenchymal and neuroectodermal cells in normal tissue, and neoplastic cells that had undergone EMT in a high proportion of canine cancers . Along with Vimentin, tumour sections and primary cell lines consistently expressed biologically active MMP2 as revealed by IF and zymography.…”

Section: Discussionsupporting

confidence: 91%

Canine oral primary melanoma cells exhibit shift to mesenchymal phenotype and phagocytic behaviour

Schmid

Brodesser

Reifinger

et al. 2019

Vet Comparative Oncology

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Canine oral malignant melanoma (COMM) is a potentially lethal cancer disease. We established primary cell lines from mostly amelanotic primary COMM and metastases and assessed lesions and derived cells for Melan A, PNL2 and CD146 expression. Then, migration and invasion of CD146‐enriched vs ‐depleted COMM cells were analysed. Epithelial‐to‐mesenchymal transition (EMT) was addressed by Vimentin‐staining and MMP2/MMP9 zymography. Phagocytic behaviour was analysed by histopathological examination and phagocytosis assay. While Melan A‐ and PNL2‐staining yielded inconsistent data, 100% of COMM sections and primary cells showed CD146 expression, suggesting that this protein may serve as a prognostic marker. An overall correlation between CD146‐expression and migration/invasion was not observed. All primary cell lines consistently expressed Vimentin and secreted biologically active MMP2, indicating that they had undergone EMT. Importantly, COMM sections exhibited cell‐in‐cell structures, and all primary cell lines exhibited phagocytic activity, supporting the concept that cell cannibalism may have a role in COMM progression.

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Section: Discussionsupporting

confidence: 91%

Canine oral primary melanoma cells exhibit shift to mesenchymal phenotype and phagocytic behaviour

Schmid

Brodesser

Reifinger

et al. 2019

Vet Comparative Oncology

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“…Since the air-dried and methanol-fixed smears are commonly used for cytology in the veterinary field, methanol was found to be the best fixative for AgNORs. In addition, availability of other fixatives was thought to be meaningful because our recent studies demonstrated that acetone and NBF were optimal fixatives for immunocytochemistry of cytokeratin/vimentin [12] and CD antigens [13], respectively. Since the smear slide used for fluorescence immunocytochemistry can be reused for other stains [12,13], the established AgNOR stain could possibly be utilized in combination with fluorescence immunocytochemistry.…”

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confidence: 99%

Argyrophilic nucleolar organizer regions staining for cytology smears in dogs and cats

et al. 2020

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The argyrophilic nucleolar organizer regions (AgNORs) are cellular proliferation markers, crucial for predicting the clinical course and aggressiveness of tumors. The purpose of this study was to establish an easy and practical AgNOR staining method in the cytology of dogs and cats. Air-dried cytological slides were prepared from dogs (n = 14) and cats (n = 12). Ethanol, acetone, and formalin were tested as fixatives for AgNOR staining. Subsequently, various methods of Romanowsky-based counterstains were tested before and after AgNOR staining. Clear and strong AgNOR spots were observed with all fixatives, and post-May-Grünwald staining was the best counterstaining method. The established method showed clear AgNOR spots even in the long-term storage samples and Romanowsky-stained ones.

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“…Complete blood count and blood chemical analyses, including C-reactive protein levels for dogs, were performed to evaluate the general conditions and concomitant diseases in all these cases. ICC was performed in accordance with the recently published method of the multiple fluorescent ICC (FICC) method [14]. The mouse monoclonal anti-human E-cadherin (clone 36, 1:100; BD Biosciences, Franklin Lakes, NJ, USA) and rabbit monoclonal anti-human vimentin (clone SP20, 1:200 dilution; Spring Bioscience, Pleasanton, CA, USA) were used as primary antibodies and incubated for 15 min at 37 °C.…”

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confidence: 99%

Immunocytochemical evaluation of epithelial–mesenchymal transition in epithelial tumors of dogs and cats

et al. 2021

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Epithelial-mesenchymal transition (EMT) plays a crucial role in metastasis of epithelial tumors; however, it is challenging to detect EMT by cytology. In the present study, EMT was visualized by fluorescenceimmunocytochemistry (FICC). Air-dried smears from epithelial tumors of dogs (n = 22) and cats (n = 9) were stained using mouse monoclonal anti-E-cadherin and rabbit monoclonal anti-vimentin antibodies.Enzymatic immunohistochemistry (IHC) revealed that 51.6 % (8/22 in dogs, 8/9 in cats) of the cases showed EMT. In dogs, FICC could detect EMT in 62.5 % (5/8) of those cases. In cats, FICC could detect EMT in 100 % (8/8) of the cases. In conclusion, the present FICC method could successfully detect EMT using conventional air-dried cytology smear slides.

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Rapid multiple immunofluorescent staining for the simultaneous detection of cytokeratin and vimentin in the cytology of canine tumors

Abstract: The RMIF method might be a useful tool for diagnostic cytology in veterinary medicine.

Cited by 8 publications

References 12 publications

Canine oral primary melanoma cells exhibit shift to mesenchymal phenotype and phagocytic behaviour

Canine oral primary melanoma cells exhibit shift to mesenchymal phenotype and phagocytic behaviour

Argyrophilic nucleolar organizer regions staining for cytology smears in dogs and cats

Immunocytochemical evaluation of epithelial–mesenchymal transition in epithelial tumors of dogs and cats

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