Ecto-5’-Nucleotidase Overexpression Reduces Tumor Growth in a Xenograph Medulloblastoma Model

Cappellari, Angélica Regina; Pillat, Micheli Mainardi; Souza, Héllio Danny Nóbrega de; Dietrich, Fabrícia; Oliveira, Francine Hehn de; Figueiró, Fabrício; Abujamra, Ana Lúcia; Roesler, Rafaël; Lecka, Joanna; Sévigny, Jean; Battastini, Ana Maria Oliveira; Ulrich, Henning

doi:10.1371/journal.pone.0140996

Cited by 21 publications

(13 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, temporal analysis of the expression in vivo reveals that the increase in CD73 by reactive astrocytes lags behind the increase in GFAP, and that the overall CD73 abundance peaks at fully developed reactive astrocytes already occupying the site of injury ( Nedeljkovic et al, 2006 ; Bjelobaba et al, 2011 ; Lavrnja et al, 2015 ). While studies in human and rat glioma and medulloblastoma cell lines in vitro also demonstrate significantly higher levels of CD73 compared to normal astrocytes ( Wink et al, 2003 ; Cappellari et al, 2012 , 2015 ; Xu et al, 2013 ), closer analysis reveals that CD73 differentiates primary tumor cell lines, which express CD73, from metastatic cell lines which do not express the enzyme ( Cappellari et al, 2012 ). Thus, we hypothesized that one of the roles of CD73 in astrocytes might be to act as a docking molecule which facilitates adhesion of non-migrating cells to the lamina, whereas mobile astrocytes, which move toward the site of injury, decrease the protein expression.…”

Section: Discussionmentioning

confidence: 97%

Unveiling the Role of Ecto-5′-Nucleotidase/CD73 in Astrocyte Migration by Using Pharmacological Tools

Adžić

Nedeljković

2018

Front. Pharmacol.

View full text Add to dashboard Cite

CD73 is a bifunctional glycosylphosphatidylinositol (GPI)-anchored membrane protein which functions as ecto-5′-nucleotidase and a membrane receptor for extracellular matrix protein (ECM). A large body of evidence demonstrates a critical involvement of altered purine metabolism and particularly, increased expression of CD73 in a number of human disorders, including cancer and immunodeficiency. Massive up-regulation of CD73 was also found in reactive astrocytes in several experimental models of human neuropathologies. In all the pathological contexts studied so far, the increased expression of CD73 has been associated with the altered ability of cells to adhere and/or migrate. Thus, we hypothesized that increased expression of CD73 in reactive astrocytes has a role in the process of astrocyte adhesion and migration. In the present study, the involvement of CD73 in astrocyte migration was investigated in the scratch wound assay (SW), using primary astrocyte culture prepared from neonatal rat cortex. The cultures were treated with one of the following pharmacological inhibitors which preferentially target individual functions of CD73: (a) α,β-methylene ADP (APCP), which inhibits the catalytic activity of CD73 (b) polyclonal anti-CD73 antibodies, which bind to the internal epitope of CD73 molecule and mask their surface exposure and (c) small interfering CD73-RNA (siCD73), which silences the expression of CD73 gene. It was concluded that approaches that reduce surface expression of CD73 increase migration velocity and promote wound closure in the scratch wound assay, while inhibition of the enzyme activity by APCP induces redistribution of CD73 molecules at the cell surface, thus indirectly affecting cell adhesion and migration. Application of anti-CD73 antibodies induces a decrease in CD73 activity and membrane expression, through CD73 molecules shedding and their release to the culture media. In addition, all applied pharmacological inhibitors differentially affect other aspects of astrocyte function in vitro, including reduced cell proliferation, altered expression of adenosine receptors and increased expression of ERK1/2. Altogether these data imply that CD73 participates in cell adhesion/migration and transmits extracellular signals through interactions with ECM.

show abstract

Section: Discussionmentioning

confidence: 97%

Unveiling the Role of Ecto-5′-Nucleotidase/CD73 in Astrocyte Migration by Using Pharmacological Tools

Adžić

Nedeljković

2018

Front. Pharmacol.

View full text Add to dashboard Cite

show abstract

“…In support of this statement, the no metastatic medulloblastoma cells, Daoy and ONS76, express high levels of functional NT5E, whereas the strongly metastatic cell line D283 does not express this ectonucleotidase [Cappellari et al, ]. Moreover, growth of induced tumors by inoculation of D283 cells in nude mice was abolishes by the knockdown of NT5E, suggesting that the accumulation of nucleosides had an inhibitory role on metastasis and tumor growth (Cappellari et al, ). Our observations in the present study in ovarian carcinoma derived SKOV‐3 cells are in agreement with these observations in medulloblastoma models.…”

Section: Discussionmentioning

confidence: 99%

Cellular Migration Ability Is Modulated by Extracellular Purines in Ovarian Carcinoma SKOV‐3 Cells

Martinez-Ramirez

Dı́az-Muñoz

Battastini

et al. 2017

J of Cellular Biochemistry

View full text Add to dashboard Cite

Extracellular nucleotides and nucleosides have emerged as important elements regulating tissue homeostasis. Acting through specific receptors, have the ability to control gene expression patterns to direct cellular fate. We observed that SKOV-3 cells express the ectonucleotidases: ectonucleotide pyrophosphatase 1 (ENPP1), ecto-5'-nucleotidase (NT5E), and liver alkaline phosphatase (ALPL). Strikingly, in pulse and chase experiments supplemented with ATP, SKOV-3 cells exhibited low catabolic efficiency in the conversion of ADP into AMP, but they were efficient in converting AMP into adenosine. Since these cells release ATP, we proposed that the conversion of ADP into AMP is a regulatory node associated with the migratory ability and the mesenchymal characteristics shown by SKOV-3 cells under basal conditions. The landscape of gene expression profiles of SKOV-3 cell cultures treated with apyrase or adenosine demonstrated similarities (e.g., decrease FGF16 transcript) and differences (e.g., the negative regulation of Wnt 2, and 10B by adenosine). Thus, in SKOV-3 we analyzed the migratory ability and the expression of epithelium to mesenchymal transition (EMT) markers in response to apyrase. Apyrase-treatment favored the epithelial-like phenotype, as revealed by the re-location of E-cadherin to the cell to cell junctions. Pharmacological approaches strongly suggested that the effect of Apyrase involved the accumulation of extracellular adenosine; this notion was strengthened when the incubation of the SKOV-3 cell with α,β-methylene ADP (CD73 inhibitor) or adenosine deaminase was sufficient to abolish the effect of apyrase on cell migration. Overall, adenosine signaling is a fine tune mechanism in the control of cell phenotype in cancer. J. Cell. Biochem. 118: 4468-4478, 2017. © 2017 Wiley Periodicals, Inc.

show abstract

“…However, CD73 has been part of a polemic argument regarding its involvement in the cancerous process. CD73 expression in ovarian carcinoma or medulloblastoma some studies was associated with a good prognosis [103,104], while in others it was associated with a poor prognosis [94]. Interestingly, Turcotte et al observed CD73 expression mainly in stromal cells with an EMT gene signature, and they proposed that ectonucleotidase is involved in tumor invasion [94].…”

Section: Adora2amentioning

confidence: 99%

Nucleotides and nucleoside signaling in the regulation of the epithelium to mesenchymal transition (EMT)

Martinez-Ramirez

Dı́az-Muñoz

Butanda-Ochoa

et al. 2016

Purinergic Signalling

View full text Add to dashboard Cite

The epithelium-mesenchymal transition (EMT) is an important process of cell plasticity, consisting in the loss of epithelial identity and the gain of mesenchymal characteristics through the coordinated activity of a highly regulated informational program. Although it was originally described in the embryonic development, an important body of information supports its role in pathology, mainly in cancerous and fibrotic processes. The purinergic system of inter-cellular communication, mainly based in ATP and adenosine acting throughout their specific receptors, has emerged as a potent regulator of the EMT in several pathological entities. In this context, cellular signaling associated to purines is opening the understanding of a new element in the complex regulatory network of this phenotypical differentiation process. In this review, we have summarized recent information about the role of ATP and adenosine in EMT, as a growing field with high therapeutic potential.Keywords P2 receptors . P1 receptors . Purinergic signaling . Cell migration . EMTThe purinergic system of intercellular communication General aspects The term purine (from: pure urine) was created more than 130 years ago by Emil Fischer after detailed analytical characterization of the uric acid molecule [1]. Purine molecular structure is the result of fusing pyrimidine and imidazole rings, and it exists as four N-H tautomeric forms [2]. Tautomerism of purine bases in DNA is one of the earliest reasons for mutations [3][4][5]. The interconversion of the different adenine or guanine tautomers produced mispairing with pyrimidines (which also show tautomeric forms) that may lead to changes in DNA sequence [6].Purine molecules appeared very early in the history of our planet, in the period known as Borganic evolution^, before the establishment of living systems. It has been postulated that purines could be formed by a eutectic (denoting a mixture of substances in fixed proportions that melts and freezes at a single temperature that is lower than the melting points of any of the separate constituents) concentration of HCN at extremely low temperature over a period of dozens of years [7]. Purine molecules, such as adenine and guanine, are components of the genetic material (DNA and RNA) of all living beings. Purine and pyrimidine tautomeric equilibria in nucleic acids was postulated to be significant in events such as DNA replication and repair as well as in the occurrence of point mutations [8].As nucleosides and nucleotides, purines play other highly strategic roles, acting as energy intermediates, allosteric regulators of key metabolic activities, redox molecules, and chemical messengers for signal transduction events. The two most important purines serving as extracellular ligands for paracrine and autocrine signaling are ATP and its dephosphorylated form, adenosine (ADO). ATP and ADO act as intracellular intermediate metabolites, and their presence in the

show abstract

Ecto-5’-Nucleotidase Overexpression Reduces Tumor Growth in a Xenograph Medulloblastoma Model

Cited by 21 publications

References 23 publications

Unveiling the Role of Ecto-5′-Nucleotidase/CD73 in Astrocyte Migration by Using Pharmacological Tools

Unveiling the Role of Ecto-5′-Nucleotidase/CD73 in Astrocyte Migration by Using Pharmacological Tools

Cellular Migration Ability Is Modulated by Extracellular Purines in Ovarian Carcinoma SKOV‐3 Cells

Nucleotides and nucleoside signaling in the regulation of the epithelium to mesenchymal transition (EMT)

Contact Info

Product

Resources

About