Systemic Administration of Small Interfering RNA Targeting Human Nestin Inhibits Pancreatic Cancer Cell Proliferation and Metastasis

Matsuda, Yoko; Ishiwata, Toshiyuki; Yoshimura, Hisashi; Yamashita, Satoshi; Arai, Tomio

doi:10.1097/mpa.0000000000000427

Cited by 21 publications

(31 citation statements)

References 31 publications

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“…In this study, we used serum-containing medium because PANC-1 cells most rapidly form large spheres under these conditions. The spheres generated from these cells expressed high mRNA levels of CSC markers, suggesting that they contain CSCs (7,13,(17)(18)(19). A previous study showed that PANC-1 cells grown in a serum-containing medium formed clusters that were heterogeneous for dye efflux, while cells grown in the serum-free medium with supplements commonly used to culture stem cells gave rise to colonies that varied in their ability to efflux dye and respond to verapamil (27).…”

Section: Discussionmentioning

confidence: 99%

“…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). Moreover, pancreatic cancer cells derived from metastatic foci of immunodeficient mice form a greater number of spheres in low-attachment plates than do their primary tumor counterparts (14).…”

Section: Introductionmentioning

confidence: 99%

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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: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2017

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“…CSCs are niche forming cells enriched with growth factors, and growing them in serum-free conditions containing growth factors, such as epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF), maintains the undifferentiated stem cell state and induces the proliferation of self-renewing, unipotent CSCs from parental cell lines [4, 25, 26]. CSCs are characterised by specific surface markers such as CD133 + /CXCR4 + , CD24 + /CD44 + , CD24 + /CD44 + /ESA + , c-Met + /CD44 + , and ALDH1 + /CD133 + in pancreatic cancer [27, 28]; CD24 −/low /CD44 + in breast cancer; CD44 + in colon/ gastric/ head and neck/ovarian cancer; CD34 + /CD38 − in leukaemia cells; CD13/CD45/CD90 in liver cancer; CD117/CD90/EpCAM in lung cancer; CD20/CD166/Nestin in melanoma cancer; and CD133 + /ABCG2 + in Glioblastoma Multiforme [29, 30]. CSCs also express various markers such as CXCR4/ ESA and Nestin [27].…”

Section: Cancer Stem Cellsmentioning

confidence: 99%

“…CSCs are characterised by specific surface markers such as CD133 + /CXCR4 + , CD24 + /CD44 + , CD24 + /CD44 + /ESA + , c-Met + /CD44 + , and ALDH1 + /CD133 + in pancreatic cancer [27, 28]; CD24 −/low /CD44 + in breast cancer; CD44 + in colon/ gastric/ head and neck/ovarian cancer; CD34 + /CD38 − in leukaemia cells; CD13/CD45/CD90 in liver cancer; CD117/CD90/EpCAM in lung cancer; CD20/CD166/Nestin in melanoma cancer; and CD133 + /ABCG2 + in Glioblastoma Multiforme [29, 30]. CSCs also express various markers such as CXCR4/ ESA and Nestin [27]. CD44 is one of the most important CSC markers for its role in promoting tumour metastasis and invasion.…”

Section: Cancer Stem Cellsmentioning

confidence: 99%

Cancer stem cell metabolism: a potential target for cancer therapy

2016

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Cancer Stem cells (CSCs) are a unipotent cell population present within the tumour cell mass. CSCs are known to be highly chemo-resistant, and in recent years, they have gained intense interest as key tumour initiating cells that may also play an integral role in tumour recurrence following chemotherapy. Cancer cells have the ability to alter their metabolism in order to fulfil bio-energetic and biosynthetic requirements. They are largely dependent on aerobic glycolysis for their energy production and also are associated with increased fatty acid synthesis and increased rates of glutamine utilisation. Emerging evidence has shown that therapeutic resistance to cancer treatment may arise due to dysregulation in glucose metabolism, fatty acid synthesis, and glutaminolysis. To propagate their lethal effects and maintain survival, tumour cells alter their metabolic requirements to ensure optimal nutrient use for their survival, evasion from host immune attack, and proliferation. It is now evident that cancer cells metabolise glutamine to grow rapidly because it provides the metabolic stimulus for required energy and precursors for synthesis of proteins, lipids, and nucleic acids. It can also regulate the activities of some of the signalling pathways that control the proliferation of cancer cells.This review describes the key metabolic pathways required by CSCs to maintain a survival advantage and highlights how a combined approach of targeting cellular metabolism in conjunction with the use of chemotherapeutic drugs may provide a promising strategy to overcome therapeutic resistance and therefore aid in cancer therapy.

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“…However, additional studies revealed that nestin was not a unique marker of neural progenitor/stem cells as the intermediate filament protein was detected in fibroblasts (e.g., heart, kidney), vascular smooth muscle and endothelial cells, pre‐existing cardiomyocytes bordering the peri‐infarct region of the ischemically damaged human/rodent heart, skeletal muscle, and tumorigenic cells (Béguin, Gosselin, Mamarbachi, & Calderone, ; Carlsson, Li, Paulin, & Thornell, ; Chen et al, ; El‐Helou et al, ; Hertig et al, ; Ishiwata et al, ; Meus, Hertig, Villeneuve, Jasmin, & Calderone, Mokry et al, ; Oikawa, Hayashi, Maesawa, Masuda, & Sobue, ; Sejersen & Lendahl, ; Tardif et al, ; Tomioka et al, ; Vaittinen et al, ). Biologically, nestin plays a direct role in the proliferation and migration of normal and tumorigenic cells (Béguin et al, ; Chen et al, ; Liang et al, ; Ishiwata, Matsuda, & Naito, ; Matsuda et al, ; Meus et al, ; Tardif et al, ; Xue & Yuan, ). Three specific regulatory intron regions identified in the nestin gene direct cell‐specific expression of the intermediate filament protein.…”

Section: Introductionmentioning

confidence: 99%

Nestin expression is dynamically regulated in cardiomyocytes during embryogenesis

Hertig

Matos‐Nieves

Garg

et al. 2017

Journal Cellular Physiology

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The transcriptional factors implicated in the expression of the intermediate filament protein nestin in cardiomyocytes during embryogenesis remain undefined. In the heart of 9,5-10,5 day embryonic mice, nestin staining was detected in atrial and ventricular cardiomyocytes and a subpopulation co-expressed Tbx5. At later stages of development, nestin immunoreactivity in cardiomyocytes gradually diminished and was absent in the heart of 17,5 day embryonic mice. In the heart of wild type 11,5 day embryonic mice, 54 ± 7% of the trabeculae expressed nestin and the percentage was significantly increased in the hearts of Tbx5 and Gata4 embryos. The cell cycle protein Ki67 and transcriptional coactivator Yap-1 were still prevalent in the nucleus of nestin -cardiomyocytes identified in the heart of Tbx5 and Gata4 embryonic mice. Phorbol 12,13-dibutyrate treatment of neonatal rat ventricular cardiomyocytes increased Yap-1 phosphorylation and co-administration of the p38 MAPK inhibitor SB203580 led to significant dephosphorylation. Antagonism of dephosphorylated Yap-1 signalling with verteporfin inhibited phorbol 12,13-dibutyrate/SB203580-mediated nestin expression and BrdU incorporation of neonatal cardiomyocytes. Nestin depletion with an AAV9 containing a shRNA directed against the intermediate filament protein significantly reduced the number of neonatal cardiomyocytes that re-entered the cell cycle. These findings demonstrate that Tbx5- and Gata4-dependent events negatively regulate nestin expression in cardiomyocytes during embryogenesis. By contrast, dephosphorylated Yap-1 acting via upregulation of the intermediate filament protein nestin plays a seminal role in the cell cycle re-entry of cardiomyocytes. Based on these data, an analogous role of Yap-1 may be prevalent in the heart of Tbx5 and Gata4 mice.

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Systemic Administration of Small Interfering RNA Targeting Human Nestin Inhibits Pancreatic Cancer Cell Proliferation and Metastasis

Abstract: Nestin plays a key role in pancreatic cancer cell metastasis and stemness and that administration of nestin siRNA may offer a novel therapeutic strategy for pancreatic cancer.

Cited by 21 publications

References 31 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

Cancer stem cell metabolism: a potential target for cancer therapy

Nestin expression is dynamically regulated in cardiomyocytes during embryogenesis

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