Enzymatic Spermine Metabolites Induce Apoptosis Associated with Increase of p53, caspase-3 and miR-34a in Both Neuroblastoma Cells, SJNKP and the N-Myc-Amplified Form IMR5
Yuta Kanamori,
Alessia Finotti,
Laura Di Magno,
Gianluca Canettieri,
Tomoaki Tahara,
Fabio Timeus,
Antonio Greco,
Paola Tirassa,
Jessica Gasparello,
Pasquale Fino,
Carlo Maria Di Liegro,
Patrizia Proia,
Gabriella Schiera,
Italia Di Liegro,
Roberto Gambari,
Enzo Agostinelli
Affiliations
Yuta Kanamori
Department of Biochemical Sciences “A. Rossi Fanelli”, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Rome, Italy
Alessia Finotti
Department of Life Sciences and Biotechnology, Biochemistry and Molecular Biology Section, University of Ferrara, 44121 Ferrara, Italy
Laura Di Magno
Department of Molecular Medicine, Sapienza University of Rome, Viale Regina Elena 291, 00161 Rome, Italy
Gianluca Canettieri
Department of Molecular Medicine, Sapienza University of Rome, Viale Regina Elena 291, 00161 Rome, Italy
Tomoaki Tahara
Department of Sensory Organs, Sapienza University of Rome, Policlinico Umberto I, Viale del Policlinico 155, 00161 Rome, Italy
Fabio Timeus
Paediatric Onco-haematology, Regina Margherita Children’s Hospital and Paediatric Department, Chivasso Hospital, 10034 Turin, Italy
Antonio Greco
Department of Sensory Organs, Sapienza University of Rome, Policlinico Umberto I, Viale del Policlinico 155, 00161 Rome, Italy
Paola Tirassa
Institute of Biochemistry and Cell Biology, Research Council of Italy (CNR), 00161 Rome, Italy
Jessica Gasparello
Department of Life Sciences and Biotechnology, Biochemistry and Molecular Biology Section, University of Ferrara, 44121 Ferrara, Italy
Pasquale Fino
UOC of Dermatology, Policlinico Umberto I Hospital, Sapienza Medical School of Rome, Viale del Policlinico 155, 00161 Rome, Italy
Carlo Maria Di Liegro
Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche) (STEBICEF), University of Palermo, 90128 Palermo, Italy
Patrizia Proia
Department of Psychology, Educational Science and Human Movement (Dipartimento di Scienze Psicologiche, Pedagogiche, dell’Esercizio fisico e della Formazione), University of Palermo, 90128 Palermo, Italy
Gabriella Schiera
Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche) (STEBICEF), University of Palermo, 90128 Palermo, Italy
Italia Di Liegro
Department of Biomedicine, Neurosciences and Advanced Diagnostics, University of Palermo, 90127 Palermo, Italy
Roberto Gambari
Department of Life Sciences and Biotechnology, Biochemistry and Molecular Biology Section, University of Ferrara, 44121 Ferrara, Italy
Enzo Agostinelli
Department of Sensory Organs, Sapienza University of Rome, Policlinico Umberto I, Viale del Policlinico 155, 00161 Rome, Italy
Neuroblastoma (NB) is a common malignant solid tumor in children and accounts for 15% of childhood cancer mortality. Amplification of the N-Myc oncogene is a well-established poor prognostic marker in NB patients and strongly correlates with higher tumor aggression and resistance to treatment. New therapies for patients with N-Myc-amplified NB need to be developed. After treating NB cells with BSAO/SPM, the detection of apoptosis was determined after annexin V-FITC labeling and DNA staining with propidium iodide. The mitochondrial membrane potential activity was checked, labeling cells with the probe JC-1 dye. We analyzed, by real-time RT-PCR, the transcript of genes involved in the apoptotic process, to determine possible down- or upregulation of mRNAs after the treatment on SJNKP and the N-Myc-amplified IMR5 cell lines with BSAO/SPM. The experiments were carried out considering the proapoptotic genes Tp53 and caspase-3. After treatment with BSAO/SPM, both cell lines displayed increased mRNA levels for all these proapoptotic genes. Western blotting analysis with PARP and caspase-3 antibody support that BSAO/SPM treatment induces high levels of apoptosis in cells. The major conclusion is that BSAO/SPM treatment leads to antiproliferative and cytotoxic activity of both NB cell lines, associated with activation of apoptosis.
