5′-nucleotidase activity in cultured cell lines. Effect of different assay conditions and correlation with cell proliferation

Turnay, Javier; Olmo, Nieves; Risse, G; K, von der Mark; Ma, Lizarbe

doi:10.1007/bf02624141

Cited by 27 publications

(10 citation statements)

References 26 publications

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“…Ecto-5'-nucleotidase is expressed or its expression is increased in cells during the process of maturation, migration or growth. This was documented for a variety of tissues and cellular systems (DePierre & Karnovsky, 1974a,b;Rodan et al, 1977;Uusitalo & Karnovsky, 1977;Dornand et al, 1986;Turnay et al, 1989) as well as for numerous tumours (Stefanovic et al, 1976;Carraway et al, 1979;Mescher et al, 1981;Amano et al, 1983;Slakey, 1985;Haffner et al, 1988;Kruger et al, 1991).…”

Section: Maturation-dependent Expressionmentioning

confidence: 88%

5′-Nucleotidase: molecular structure and functional aspects

Zimmermann

1992

Biochemical Journal

806

701

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Section: Maturation-dependent Expressionmentioning

confidence: 88%

5′-Nucleotidase: molecular structure and functional aspects

Zimmermann

1992

Biochemical Journal

806

701

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“…It has been demonstrated that cells subjected to scratch wound increase cell proliferation ( Lampugnani, 1999 ; Kornyei et al, 2000 ), whereas the cell proliferation positively correlates with CD73 expression in many different tumor cell lines, including glioma ( Turnay et al, 1989 ; Christensen et al, 1996 ; Ciccarelli et al, 2000 ; Ohana et al, 2001 ; Bavaresco et al, 2008 ; Gao et al, 2017 ). Cell proliferation and CD73 expression are interconnected through TCF/LEF binding site in the regulatory region of CD73 gene, which is the nuclear target of Wnt pathway ( Synnestvedt et al, 2002 ; Spychala and Kitajewski, 2004 ).…”

Section: Discussionmentioning

confidence: 99%

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

Adžić

Nedeljković

2018

Front. Pharmacol.

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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.

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“…It is interesting to note that adenosine itself stimulates 5hnucleotidase in rat mesangial cells [39]. In different cells its activity increases during the exponential growth phase of the cells in culture and is significantly down-regulated once the cells reach confluency [40].…”

Section: Discussionmentioning

confidence: 99%

Regulation of proliferation of LLC-MK2 cells by nucleosides and nucleotides: the role of ecto-enzymes

Lemmens

Vanduffel

Teuchy

et al. 1996

Biochemical Journal

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1. Using the incorporation of [methyl-3H]thymidine as a proliferation marker, the effects of various nucleosides and nucleotides on endothelial LLC-MK2 cells were studied. We found that ATP, ADP, AMP and adenosine in concentrations of 10 microM or higher stimulate the proliferation of these cells. 2. Inhibition of ecto-ATPase (EC 3.6.1.15), 5'-nucleotidase (EC 3.1.3.5) or alkaline phosphatase (EC 3.1.3.1) significantly diminished the stimulatory effect of ATP, indicating that the effect is primarily caused by adenosine and not by adenine nucleotides. Also, the effect depends only on extracellular nucleosides, since inhibition of nucleoside uptake by dipyridamole has no influence on proliferation. 3. Other purine nucleotides and nucleosides (ITP, GTP, inosine and guanosine) also stimulate cell proliferation, while pyrimidine nucleotides and nucleosides (CTP, UTP, cytidine and uridine) inhibit proliferation. Furthermore, the simultaneous presence of adenosine and any of the other purine nucleosides is not entirely additive in its effect on cell proliferation. At the same time any pyrimidine nucleoside, when added together with adenosine, has the same inhibitory effect as the pyrimidine nucleoside alone. 4. Apparently these proliferative effects are neither caused by any pharmacologically known P1-purinoceptor, nor are they mediated by cyclic AMP, cyclic GMP, or D-myo-inositol 1,4,5-trisphosphate as second messenger, nor by extracellular Ca2+. 5. Therefore, we conclude that various purine and pyrimidine nucleosides can influence the proliferation of LLC-MK2 cells by acting on putative purinergic and pyrimidinergic receptors not previously described.

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5′-nucleotidase activity in cultured cell lines. Effect of different assay conditions and correlation with cell proliferation

Cited by 27 publications

References 26 publications

5′-Nucleotidase: molecular structure and functional aspects

5′-Nucleotidase: molecular structure and functional aspects

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

Regulation of proliferation of LLC-MK2 cells by nucleosides and nucleotides: the role of ecto-enzymes

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