title: Mitochondrial fusion: a mechanism of cisplatin-induced resistance in neuroblastoma cells?
abstract: Cisplatin induces apoptosis through different pathways. The intrinsic apoptotic pathway is mediated by mitochondria, which, as a result of cisplatin treatment, undergo morphological alterations. The aim of this study was to investigate cisplatin-induced mitochondrial functional and morphological long-term effects in neuroblastoma B50 rat cells. To this purpose, we followed evaluated different several apoptotic markers by means of flow cytometry, confocal and electron microscopy and western blotting techniques. We applied different treatment protocols based on the incubation of the neuroblastoma B50 rat cells with 40 μM cisplatin: (i) for 48 h and harvesting of the cells at the end of the treatment; (ii) further recovery in drug-free medium for 7 days post-treatment; (iii) conditions as in (ii) followed by re-seeding in normal medium and growth for a further 4 days. We observed apoptosis induction after the first treatment and after the recovery from cell death after long-term culture in drug-free medium. Interestingly, the latter phenomenon was characterized by mitochondrial elongation and mitochondrial protein rearrangement. In recovered and re-seeded cells, mitochondrial equilibrium moved toward fusion, possibly protecting cells from apoptosis.
authors.affiliation: Istituto di Genetica Molecolare del CNR, Università di Pavia, via Ferrata 9, 27100 Pavia, Italy. giada.santin@unipv.it
authors.forename: Giada
authors.initials: G
authors.lastname: Santin
authors.forename: Valeria M
authors.initials: VM
authors.lastname: Piccolini
authors.forename: Sergio
authors.initials: S
authors.lastname: Barni
authors.forename: Paola
authors.initials: P
authors.lastname: Veneroni
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Giansanti
authors.forename: Veronica
authors.initials: V
authors.lastname: Dal Bo
authors.forename: Graziella
authors.initials: G
authors.lastname: Bernocchi
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Bottone
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D045304:Cytochromes c / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D020558:GTP Phosphohydrolases / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D063154:Mitochondrial Dynamics / Q000187:drug effects*; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D056564:Sirtuin 1 / Q000378:metabolism; D013997:Time Factors; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000199:Actins; C494831:Aifm1 protein, rat; D000970:Antineoplastic Agents; D051033:Apoptosis Inducing Factor; C494828:Bax protein, rat; D004272:DNA, Mitochondrial; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; C499322:Parp1 protein, rat; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C488368:Sirt1 protein, rat; D056564:Sirtuin 1; D020558:GTP Phosphohydrolases; C498811:Opa1 protein, rat; D002945:Cisplatin