title: Cell proliferation and drug sensitivity of human glioblastoma cells are altered by the stable modulation of cytosolic 5'-nucleotidase II.
abstract: Cytosolic 5'-nucleotidase II (cN-II) has been reported to be involved in cell survival, nucleotide metabolism and in the cellular response to anticancer drugs. With the aim to further evaluate the role of this enzyme in cell biology, we stably modulated its expression the human glioblastoma cell ADF in which the transient inhibition of cN-II has been shown to induce cell death. Stable cell lines were obtained both with inhibition, obtained with plasmids coding cN-II-targeting short hairpin RNA, and stimulation, obtained with plasmids coding Green Fluorescence Protein (GFP)-fused wild type cN-II or a GFP-fused hyperactive mutant (GFP-cN-II-R367Q), of cN-II expression. Silenced cells displayed a decreased proliferation rate while the over expressing cell lines displayed an increased proliferation rate as evidenced by impedance measurement using the xCELLigence device. The expression of nucleotide metabolism relevant genes was only slightly different between cell lines, suggesting a compensatory mechanism in transfected cells. Cells with decreased cN-II expression were resistant to the nucleoside analog fludarabine confirming the involvement of cN-II in the metabolism of this drug. Finally, we observed sensitivity to cisplatin in cN-II silenced cells and resistance to this same drug in cN-II over-expressing cells indicating an involvement of cN-II in the mechanism of action of platinum derivatives, and most probably in DNA repair. In summary, our findings confirm some previous data on the role of cN-II in the sensitivity of cancer cells to cancer drugs, and suggest its involvement in other cellular phenomenon such as cell proliferation.
authors.affiliation: University of Pisa, Department of Biology, Biochemistry Unit, Pisa, Italy. Electronic address: f.cividini@gmail.com.
authors.forename: F
authors.initials: F
authors.lastname: Cividini
authors.affiliation: Université de Lyon, F-69000 Lyon, France; Université de Lyon 1, F-69622 Lyon, France; Université de Lyon 1, F-69000 Lyon, France; INSERM U1052, Centre de Recherche en Cancérologie de Lyon, F-69000 Lyon, France; CNRS UMR 5286, Centre de Recherche en Cancérologie de Lyon, F-69000 Lyon, France.
authors.forename: E
authors.initials: E
authors.lastname: Cros-Perrial
authors.affiliation: University of Pisa, Department of Biology, Biochemistry Unit, Pisa, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Pesi
authors.affiliation: Hospices Civils de Lyon, Centre Hospitalier Lyon-Sud, Laboratoire de Biochimie et Toxicologie, Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Machon
authors.affiliation: University of Sassari, Department of Chemistry and Pharmacology, Sassari, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Allegrini
authors.affiliation: University of Pisa, Department of Biology, Biochemistry Unit, Pisa, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Camici
authors.affiliation: Université de Lyon, F-69000 Lyon, France; Université de Lyon 1, F-69622 Lyon, France; Université de Lyon 1, F-69000 Lyon, France; INSERM U1052, Centre de Recherche en Cancérologie de Lyon, F-69000 Lyon, France; CNRS UMR 5286, Centre de Recherche en Cancérologie de Lyon, F-69000 Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Dumontet
authors.affiliation: Université de Lyon, F-69000 Lyon, France; Université de Lyon 1, F-69622 Lyon, France; Université de Lyon 1, F-69000 Lyon, France; INSERM U1052, Centre de Recherche en Cancérologie de Lyon, F-69000 Lyon, France; CNRS UMR 5286, Centre de Recherche en Cancérologie de Lyon, F-69000 Lyon, France.
authors.forename: L P
authors.initials: LP
authors.lastname: Jordheim
authors.affiliation: University of Pisa, Department of Biology, Biochemistry Unit, Pisa, Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Tozzi
mesh_terms: D015720:5'-Nucleotidase / Q000235:genetics / Q000378:metabolism*; D049109:Cell Proliferation / Q000502:physiology; D055785:Gene Knockdown Techniques; D005909:Glioblastoma / Q000188:drug therapy* / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D006801:Humans; D014162:Transfection
chemical_list: D015720:5'-Nucleotidase; C508048:NT5C2 protein, human
keywords: Anticancer drugs; Cell proliferation; Cytosolic 5′-nucleotidase II; Human glioblastoma ADF cells; Nucleotide metabolism; Stable protein modulation