Isolamento, cultura e efeito de Viscum album L., doxorrubicina e associação na morfologia e expressão proteica de linhagem celular primária de adenomioepitelioma maligno canino
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Data
2025-04-25
Autores
Angelos, Tamires de Almeida
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Universidade Federal do Espírito Santo
Resumo
Breast cancer is the disease that most affects female dogs in veterinary medicine, lacking precise therapies that affect only cancer cells, helping in tumor eradication and survival. Thus, the objective of this study was to isolate and cultivate a primary canine malignant adenomyoepithelioma (AME) cell line, and to evaluate the individual effects and the association of Viscum album L. (D3 dilution) and doxorubicin on the morphology and expression of BAX, BCL-XL, OGG, GPX and REF1 of a primary canine malignant adenomyoepithelioma cell line. A canine malignant adenomyoepithelioma sample from a 9-year-old SRD bitch was used for isolation and primary culture of cancer cells. Previously, the working LC50 of Viscum album L. (D3 dilution) was established in 24 and 48h (37.07 μL/mL), as well as the working concentration of doxorubicin, also after 24 and 48h of viability assay, defining the concentration 1.56 μg/mL, using cancer cells in primary culture, 20th pass. The concentrations determined were used for cell morphology and protein expression assays by immunohistochemistry after exposure for 24 and 48 hours to the treatments Viscum album L., doxorubicin and Viscum album L + doxorubicin association. The morphological findings showed minimal morphological differences compared to the control group, acting mainly as a cytoprotective agent, while doxorubicin alone and in association strongly induced cell death. The weak positive expression score maintained at 24 hours in the BAX, BCL-XL and GPX proteins in the Viscum album L. and doxorubicin groups compared to the control group, differing only from the associated treatment Viscum album L. + doxorubicin. At 48 hours, the weak positive score remained present in all treatments compared to the control group in GPX and BAX, differing only in BCL-XL the Viscum album L. and doxorubicin groups compared to the control and association group that presented a positive score. When comparing the times, it was obtained that doxorubicin increases the expression of GPX and BAX in addition to decreasing BCL-XL in 48 hours while the association acts by increasing the pro apoptotic protein. Viscum album L. decreased the nuclear labeling of OGG, while doxorubicin increased OGG and decreased the labeling of REF-1. The association of the compounds led to an increase in OGG and a decrease in REF-1. It is mainly concluded that the association of Viscum album L. and doxorubicin potentiate their mechanisms of action in a time-response manner, in addition to the fact that in SMA, isolated treatments induce the activation of other types of programmed cell death, in addition to apoptosis.
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Cultivo primário , Estresse oxidativo , Oncologia veterinária