title: Profiling the molecular mechanism of fullerene cytotoxicity on tumor cells by RNA-seq.
abstract: The interest on functionalized fullerenes in the field of nanomedicine has seen a significant increase in the past decade. However, the different methods employed to increase C60 solubility profoundly influence the physicochemical properties and the toxicological effects of these compounds, thus complicating the evaluation of their toxicity and potential therapeutic use. Here we report a whole-transcriptome RNA-seq analysis assessing the effect of two fullerenes (1 and 2) on gene expression in the human MCF7 cell line. Although these two compounds had previously been characterized by in vitro studies as having a cytotoxic and null effect respectively, to date the mechanisms at the basis of this different behavior and, more in general, at the basis of the effect of most fullerene derivatives in living cells are still completely unknown. Our data evidence that: (a) fullerene 2 caused a significant, time-dependent alteration of gene expression, whereas 1 only had a negligible effect; (b) the biological processes mostly influenced over the 48h experimental time course were transcription, protein synthesis, cell cycle progression and cell adhesion; (c) the gene expression signature of 2-treated cells was strikingly similar to those induced by selective inhibitors of mTOR signaling, thus suggesting an effect on this pathway for fullerene 2. Our work represents the first approach toward the application of RNA-seq to the study of the molecular mechanisms underlying the interaction of fullerenes with cellular systems and provides an objective view of the feasibility and the safety of these nanomaterials for a medical application.
authors.affiliation: Department of Life Sciences, University of Trieste, Via Giorgieri 5, 34127 Trieste (TS), Italy. Electronic address: marianna.lucafo@phd.units.it.
authors.forename: Marianna
authors.initials: M
authors.lastname: Lucafò
authors.forename: Marco
authors.initials: M
authors.lastname: Gerdol
authors.forename: Alberto
authors.initials: A
authors.lastname: Pallavicini
authors.forename: Sabrina
authors.initials: S
authors.lastname: Pacor
authors.forename: Sonia
authors.initials: S
authors.lastname: Zorzet
authors.forename: Tatiana
authors.initials: T
authors.lastname: Da Ros
authors.forename: Maurizio
authors.initials: M
authors.lastname: Prato
authors.forename: Gianni
authors.initials: G
authors.lastname: Sava
mesh_terms: D000970:Antineoplastic Agents; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D005109:Extracellular Matrix / Q000187:drug effects; D037741:Fullerenes / Q000737:chemistry / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D049329:Nanostructures / Q000633:toxicity; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000187:drug effects / Q000235:genetics*; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000187:drug effects; D059467:Transcriptome / Q000187:drug effects; D016212:Transforming Growth Factor beta / Q000187:drug effects / Q000502:physiology; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D037741:Fullerenes; D009363:Neoplasm Proteins; D016212:Transforming Growth Factor beta; D012313:RNA; D004247:DNA; D058570:TOR Serine-Threonine Kinases
keywords: Fullerene; Gene expression; Nanotoxicology; Next generation sequencing; RNA-seq