Dataset Information

Matrine Targets BTF3 to Inhibit the Growth of Canine Mammary Tumor Cells.

ABSTRACT: The canine mammary tumor model is more suitable for studying human breast cancer, and the safety concentrations of matrine and the biotin-labeled matrine probe were determined in canine primary mammary epithelial cells, and then selected canine mammary tumor cell lines CHMm and CHMp were incubated with matrine, and cell viability was detected by CCK-8. The biotin-labeled matrine probe was used to pull-down the targets of matrine in canine mammary tumor cells, and the targets were screened in combination with activity-based protein profiling (ABPP) and Genecards database, and verified by qPCR and western blot. The results showed that the maximum non-cytotoxic concentrations of matrine and biotin-labeled matrine probe in canine primary mammary epithelial cells were 250 μg/mL and 500 μg/mL, respectively. Matrine and biotin-labeled matrine probe had a proliferation inhibitory effect time-dependently on CHMm and CHMp cells within a safe concentration range, and induced autophagy in cells. Then BTF3 targets were obtained by applying ABPP and Genecards screening. Cellular thermal shift assay (CETSA) findings indicated that matrine could increase the heat stability of BTF3 protein. Pull-down employing biotin-labeled matrine probe with CHMm and CHMp cell lysates revealed that BTF3 protein was detected in the biotin-labeled matrine probe group and that BTF3 protein was significantly decreased by the addition of matrine. The qPCR and western blot findings of CHMm and CHMp cells treated with matrine revealed that matrine decreased the expression of the BTF3 gene and protein with the extension of the action time, and the impact was more substantial at the protein level, respectively.

SUBMITTER: Feng Z

PROVIDER: S-EPMC10779273 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Matrine Targets BTF3 to Inhibit the Growth of Canine Mammary Tumor Cells.

Feng Zijian Z Sun Na N Noor Fida F Sun Panpan P Zhang Hua H Zhong Jia J Yin Wei W Fan Kuohai K Yang Huizhen H Zhang Zhenbiao Z Sun Yaogui Y Li Hongquan H

International journal of molecular sciences 20231230 1

The canine mammary tumor model is more suitable for studying human breast cancer, and the safety concentrations of matrine and the biotin-labeled matrine probe were determined in canine primary mammary epithelial cells, and then selected canine mammary tumor cell lines CHMm and CHMp were incubated with matrine, and cell viability was detected by CCK-8. The biotin-labeled matrine probe was used to pull-down the targets of matrine in canine mammary tumor cells, and the targets were screened in com ...[more]

PMID: 38203709

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Project description:BackgroundCancer spread to other organs is the main cause of death of oncological patients. Migration of cancer cells from a primary tumour is the crucial step in the complex process of metastasis, therefore blocking this process is currently the main treatment strategy. Metastasis inhibitors derived from natural products, such as, migrastatin, are very promising anticancer agents. Thus, the aim of our study was to investigate the effect of six migrastatin analogues (MGSTA-1 to 6) on migration and invasion of canine mammary adenocarcinoma cell lines isolated from primary tumours and their metastases to the lungs. Canine mammary tumours constitute a valuable tool for studying multiple aspect of human cancer.ResultsOUR RESULTS SHOWED THAT TWO OF SIX FULLY SYNTHETIC ANALOGUES OF MIGRASTATIN: MGSTA-5 and MGSTA-6 were potent inhibitors of canine mammary cancer cells migration and invasion. These data were obtained using the wound healing test, as well as trans-well migration and invasion assays. Furthermore, the treatment of cancer cells with the most effective compound (MGSTA-6) disturbed binding between filamentous F-actin and fascin1. Confocal microscopy analyses revealed that treatment with MGSTA-6 increased the presence of unbound fascin1 and reduced co-localization of F-actin and fascin1 in canine cancer cells. Most likely, actin filaments were not cross-linked by fascin1 and did not generate the typical filopodial architecture of actin filaments in response to the activity of MGSTA-6. Thus, administration of MGSTA-6 results in decreased formation of filopodia protrusions and stress fibres in canine mammary cancer cells, causing inhibition of cancer migration and invasion.ConclusionTwo synthetic migrastatin analogues (MGSTA-5 and MGSTA-6) were shown to be promising compounds for inhibition of cancer metastasis. They may have beneficial therapeutic effects in cancer therapy in dogs, especially in combination with other anticancer drugs. However, further in vivo studies are required to verify this hypothesis.

Project description:IntroductionIntestinal lymphoma may be latent in some dogs with chronic inflammatory enteropathy. Mesenchymal stromal cells (MSCs) have potential therapeutic applications for refractory chronic inflammatory enteropathy, but their impact on the development of potential intestinal lymphomas has not yet been evaluated. Therefore, this study was performed to investigate the effect of canine adipose-derived MSCs (cADSCs) on the growth of canine lymphoma cell lines to assess the safety of MSC-based therapy in terms of pro- and anti-tumorigenic effects.MethodsCADSCs were co-cultured with canine lymphoma/leukemia cell lines of various lineages, with or without cell-to-cell contact, to evaluate their effects on proliferation, apoptosis, and cell cycle progression in vitro. Additionally, a bioluminescent canine lymphoma cell line, established through firefly luciferase transduction, was co-injected with varying doses of cADSCs into immunocompromised mice. The growth of canine lymphoma cells was monitored over time in vivo using bioluminescence imaging.ResultsCADSCs inhibited the proliferation of all canine lymphoma/leukemia cell lines in a dose-dependent manner in vitro, under conditions allowing cell-to-cell contact. This inhibition occurred via the induction of apoptosis, G0/G1 phase cell cycle arrest, or both mechanisms. However, these effects were lost when the cells were physically separated using Transwell inserts. In xenotransplantation mouse models, cADSCs dose-dependently inhibited canine lymphoma cell growth and lung metastasis, as indicated by reduced bioluminescence signals.ConclusionsThis study has demonstrated for the first time that cADSCs inhibit the growth of different lineages of canine lymphoma/leukemia cells both in vitro and in vivo. These findings suggest that MSC-based cell therapy could potentially be applied to canine chronic inflammatory enteropathy without increasing the risk of promoting the growth of latent intestinal lymphomas.

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Matrine Targets BTF3 to Inhibit the Growth of Canine Mammary Tumor Cells.

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Matrine Targets BTF3 to Inhibit the Growth of Canine Mammary Tumor Cells.

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