Characterization of spheres derived from canine mammary gland adenocarcinoma cell lines

Michishita, Masaki; Akiyoshi, R.; Yoshimura, Hisashi; Katsumoto, Takuo; Ichikawa, Hitoshi; Ohkusu-Tsukada, Kozo; Nakagawa, Takayuki; Sasaki, Nobuo; Takahashi, Koji

doi:10.1016/j.rvsc.2010.11.016

Cited by 51 publications

(84 citation statements)

References 23 publications

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“…In the latter assay, the CSCs of PDAC cells form floating colonies when cultivated in ultra-low-attachment dishes (9)(10)(11) (12). In a previous study, we reported that nestin, a pancreatic CSC marker, is more highly expressed in the spheres of three PDAC cell lines than in nonsphere cells (13).…”

Section: Introductionmentioning

confidence: 99%

Electron microscopic analysis of different cell types in human pancreatic cancer spheres

et al. 2017

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Abstract. Cancer stem cells (CSCs), which are pluripotent and self-renewable, contribute to the initiation and metastasis of cancer, and are responsible for resistance to chemotherapy and radiation. Pancreatic ductal adenocarcinoma (PDAC) is an aggressive type of cancer that is associated with a high incidence of distant metastasis and recurrence. Sphere formation reveals cell proliferation under nonadherent conditions and is commonly used to identify CSCs; measurements of the number, area and volume of the spheres are used to estimate stemness of PDAC cells. However, detailed morphological analysis of such spheres has not been performed. The aim of the present study was to examine the morphology of spheres isolated from PANC-1 human pancreatic cancer cells via scanning electron microscopy (SEM) and transmission electron microscopy (TEM). PANC-1 cells formed round to irregular oblong spheres within 1 week following seeding in ultra-low-attachment plates. These spheres exhibited higher levels of expression of CSC markers, including nestin, sex determining region Y-box 2, and CD44 containing variant exon 9, compared with adherent cells. SEM analysis revealed that the spheres exhibited a grape-like appearance, harboring cancer cells with smooth or rough surfaces. Similarly, TEM analysis detected cancer cells with varying surface types within the spheres: Those with smooth surfaces, irregular large protrusions, protrusions and a small number of microvilli, and those with many microvilli throughout the entire cell surface. These morphological differences among cancer cells may be indicative of different stages in the differentiation process, from CSCs to non-CSCs, within the spheres.

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Section: Introductionmentioning

confidence: 99%

Electron microscopic analysis of different cell types in human pancreatic cancer spheres

et al. 2017

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“…Furthermore, activation of the Wnt/β-catenin, Hedgehog and Notch signaling pathways confers resistance and survival to CSCs in human cancer (13). Similar to what has been observed in human cancer, canine mammary CSCs upregulate stem cell-associated genes (9) and activate the Wnt/β-catenin signaling pathway (14). Canine mammary CSCs are highly tumorigenic, and exhibit multidrug resistance and invasive potential (9,15); therefore, they serve an important role in the treatment resistance, recurrence and metastasis of canine mammary tumors.…”

Section: Introductionmentioning

confidence: 53%

“…Cell spheres were obtained as previously described (9). CMT7364 and CIPp cells and their spheres were plated on 96-well plates at a density of Sphere formation assay to identify CSCs.…”

Section: Cellmentioning

confidence: 99%

“…Increasing evidence supports the presence of CSCs in canine mammary tumors (7)(8)(9)(10). Human breast CSCs are primarily characterized by cluster of differentiation (CD)44 + /CD24 -/low cells, sphere formation and active aldehyde dehydrogenase (11).…”

Section: Introductionmentioning

confidence: 99%

“…-/low phenotype is also a specific maker of CSCs in canine mammary carcinoma (8,9,12). Another feature of CSCs is overexpression of stem cell-associated genes, including octamer-binding transcription factor 4 (Oct-4) and sex determining region Y-box 2 (Sox-2) (10).…”

Section: Introductionmentioning

confidence: 99%

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Salinomycin inhibits canine mammary carcinoma in vitro by targeting cancer stem cells

Zhou

Zhang

et al. 2017

Oncology Letters

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Abstract. Salinomycin (SAL), a polyether ionophore antibiotic, has been demonstrated to selectively kill cancer stem cells (CSCs) in various types of human tumor. The aim of the present study was to investigate the effects of SAL on canine mammary CSCs. CSCs in canine mammary carcinoma cell lines (CMT7364 and CIPp) were identified using a sphere formation assay and flow cytometry. The chemoresistance, invasive potential and expression of stem cell-associated proteins of these spheres was then analyzed. This demonstrated that the spheres exhibited characteristics of CSCs, including a cluster of differentiation (CD)44 + /CD24 -/low phenotype, upregulation of Wnt/β-catenin signaling pathway-associated proteins and chemoresistance. The viability of the spheres was decreased in a concentration-and time-dependent manner following treatment with SAL, and the spheres did not exhibit increased resistance to SAL compared with their parental cells. In addition, exposure to SAL inhibited sphere-formation and invasive potential in canine mammary CSCs in a dose-dependent manner. Furthermore, SAL decreased the CD44 + /CD24 -/low population and downregulated the expression of Wnt/β-catenin signaling-associated proteins (β-catenin, Cyclin D1 and octamer-binding transcription factor 4) in the spheres. In conclusion, the present study demonstrated that SAL is an effective inhibitor of canine mammary CSCs in vitro, indicating that SAL is a promising chemotherapeutic agent for the treatment of canine mammary carcinoma.

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Conserved and variable: Understanding mammary stem cells across species

2017

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Postnatal mammary gland development requires the presence of mammary stem and progenitor cells (MaSC), which give rise to functional milk-secreting cells and regenerate the mammary epithelium with each cycle of lactation. These long-lived, tissue-resident MaSC are also targets for malignant transformation and may be cancer cells-oforigin. Consequently, MaSC are extensively researched in relation to their role and function in development, tissue regeneration, lactation, and breast cancer. The basic structure and function of the mammary gland are conserved among all mammalian species, from the most primitive to the most evolved. However, species vary greatly in their lactation strategies and mammary cancer incidence, making MaSC an interesting focus for comparative research. MaSC have been characterized in mice, to a lesser degree in humans, and to an even lesser degree in few additional mammals. They remain uncharacterized in most mammalian species, including "ancient" monotremes, marsupials, wild, and rare species, as well as in common and domestic species such as cats. Identification and comparison of MaSC across a large variety of species, particularly those with extreme lactational adaptations or low mammary cancer incidence, is expected to deepen our understanding of development and malignancy in the mammary gland. Here, we review the current status of MaSC characterization across species, and underline species variations in lactation and mammary cancer through which we may learn about the role of MaSC in these processes. V C 2017 International Society for Advancement of Cytometry

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Characterization of spheres derived from canine mammary gland adenocarcinoma cell lines

Cited by 51 publications

References 23 publications

Electron microscopic analysis of different cell types in human pancreatic cancer spheres

Electron microscopic analysis of different cell types in human pancreatic cancer spheres

Salinomycin inhibits canine mammary carcinoma in vitro by targeting cancer stem cells

Conserved and variable: Understanding mammary stem cells across species

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