Decreased activity of inducible nitric oxide synthase type 2 and modulation of the expression of glutathione S-transferase alpha, bcl-2, and metallothioneins during the differentiation of CaCo-2 cells.

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State: Public
Version: Final published version
Serval ID
serval:BIB_724D1DA74EF4
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Decreased activity of inducible nitric oxide synthase type 2 and modulation of the expression of glutathione S-transferase alpha, bcl-2, and metallothioneins during the differentiation of CaCo-2 cells.
Journal
Cell Growth and Differentiation
Author(s)
Vecchini F., Pringault E., Billiar T.R., Geller D.A., Hausel P., Felley-Bosco E.
ISSN
1044-9523 (Print)
ISSN-L
1044-9523
Publication state
Published
Issued date
1997
Peer-reviewed
Oui
Volume
8
Number
2
Pages
261-268
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Reactive oxygen species modulate the cell growth of a wide variety of mammalian cells. To determine whether oxidative metabolism is altered during the differentiation process, we studied the expression of pro- and antioxidant proteins in proliferating and differentiated CaCo-2 cells, a human colon adenocarcinoma cell line. Nitric oxide synthase type 2 (iNOS) produces nitric oxide (NO). Depending on its rate of synthesis, NO may either promote cellular and DNA damage or reduce the ability of other free radicals to induce cell injury. Using Western and Northern blot analysis and arginine conversion assay, we demonstrate that the expression of iNOS decreases when cells undergo differentiation. This biological event entails a diminished production of NO metabolites and correlates with the loss of activation of soluble guanylate cyclase activity. In differentiated cells, a 2-fold down-regulation of the nuclear factor kappa B activity was observed, suggesting that nuclear factor kappa B could be one of the iNOS gene regulatory factors in the CaCo-2 model. In parallel, we studied the expression of other antioxidant proteins including glutathione S-transferase alpha (GST alpha), bcl-2, and the metallothioneins (MTs). We show that the protein levels of GST alpha and MT increase during the differentiation of CaCo-2 cells, whereas bcl-2 levels decrease. Our investigation indicates that the expression of iNOS, GST alpha, bcl-2, and MT is associated with the enterocytic differentiation. The shift in the expression of specific antioxidant genes during CaCo-2 cell differentiation may occur to avoid alterations in the cell redox potential.
Keywords
Actins/chemistry, Blotting, Western, Caco-2 Cells, Cell Differentiation/genetics, Cell Differentiation/physiology, Cyclic GMP/chemistry, Enzyme Induction, Gene Expression Regulation/physiology, Glutathione Transferase/biosynthesis, Glutathione Transferase/genetics, Humans, Intestines/cytology, Metallothionein/biosynthesis, Metallothionein/genetics, Models, Biological, Nitric Oxide Synthase/biosynthesis, Nitric Oxide Synthase/genetics, Proto-Oncogene Proteins c-bcl-2/biosynthesis, Tumor Suppressor Protein p53/chemistry
Pubmed
Web of science
Create date
03/08/2014 15:17
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20/08/2019 15:30
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