title: An intrinsic DFF40/CAD endonuclease deficiency impairs oligonucleosomal DNA hydrolysis during caspase-dependent cell death: a common trait in human glioblastoma cells.
abstract: Glioblastoma (GBM) or grade IV astrocytoma is one of the most devastating human cancers. The loss of DFF40/CAD, the key endonuclease that triggers oligonucleosomal DNA fragmentation during apoptosis, has been linked to genomic instability and cell survival after radiation. Despite the near inevitability of GBM tumor recurrence after treatment, the relationship between DFF40/CAD and GBM remains unexplored.
  
  
  
  We studied the apoptotic behavior of human GBM-derived cells after apoptotic insult. We analyzed caspase activation and the protein levels and subcellular localization of DFF40/CAD apoptotic endonuclease. DFF40/CAD was also evaluated in histological sections from astrocytic tumors and nontumoral human brain.
  
  
  
  We showed that GBM cells undergo incomplete apoptosis without generating oligonucleosomal DNA degradation despite the correct activation of executioner caspases. The major defect of GBM cells relied on the improper accumulation of DFF40/CAD at the nucleoplasmic subcellular compartment. Supporting this finding, DFF40/CAD overexpression allowed GBM cells to display oligonucleosomal DNA degradation after apoptotic challenge. Moreover, the analysis of histological slices from astrocytic tumors showed that DFF40/CAD immunoreactivity in tumoral GFAP-positive cells was markedly reduced when compared with nontumoral samples.
  
  
  
  Our data highlight the low expression levels of DFF40/CAD and the absence of DNA laddering as common molecular traits in GBM. These findings could be of major importance for understanding the malignant behavior of remaining tumor cells after radiochemotherapy.
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: María
authors.initials: M
authors.lastname: Sánchez-Osuna
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Laura
authors.initials: L
authors.lastname: Martínez-Escardó
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Carla
authors.initials: C
authors.lastname: Granados-Colomina
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Fina
authors.initials: F
authors.lastname: Martínez-Soler
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Sònia
authors.initials: S
authors.lastname: Pascual-Guiral
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Victoria
authors.initials: V
authors.lastname: Iglesias-Guimarais
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Roser
authors.initials: R
authors.lastname: Velasco
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Gerard
authors.initials: G
authors.lastname: Plans
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Noemi
authors.initials: N
authors.lastname: Vidal
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Avelina
authors.initials: A
authors.lastname: Tortosa
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Carlos
authors.initials: C
authors.lastname: Barcia
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Jordi
authors.initials: J
authors.lastname: Bruna
authors.affiliation: Cell Death, Senescence and Survival group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (M.S.-O., L.M.-E., C.G.-C., S.P.-G., V.I.-G., V.J.Y.); Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (C.I.B.E.R.N.E.D.), Barcelona, Spain (M.S.-O., V.I.-G., V.J.Y.); Department of Basic Nursing, Institut d'Investigació Biomèdica de Bellvitge-Universitat de Barcelona, Barcelona, Spain (F.M.-S., A.T.); Unit of Neuro-Oncology, Hospital Universitari de Bellvitge-I.C.O Duran i Reynals, Barcelona, Spain (R.V., G.P., N.V., J.B.); Group of Neuroplasticity and Regeneration (C.I.B.E.R.N.E.D.), Department of Cell Biology, Physiology and Immunology & Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain (R.V., J.B.); Neuropathology Institute, Hospital Universitari de Bellvitge, Barcelona, Spain (N.V.); Neuro-Immunity Group, Department of Biochemistry and Molecular Biology and Institute of Neurosciences, Faculty of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain (C.B.).
authors.forename: Victor J
authors.initials: VJ
authors.lastname: Yuste
mesh_terms: D017209:Apoptosis / Q000235:genetics* / Q000502:physiology; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D020169:Caspases / Q000378:metabolism*; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000172:deficiency*; D005095:Exoribonucleases / Q000235:genetics*; D005909:Glioblastoma / Q000201:enzymology*; D006801:Humans; D000075223:Poly-ADP-Ribose Binding Proteins
chemical_list: D051017:Apoptosis Regulatory Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D004247:DNA; C113696:DFFB protein, human; D003851:Deoxyribonucleases; D005095:Exoribonucleases; C065486:oligonucleotidase; D020169:Caspases
keywords: apoptosis; caspase-dependent DNase (DFF40/CAD); caspase-dependent cell death; glioblastoma; oligonucleosomal DNA degradation