title: Evaluating polyethyleneimine/DNA nanoparticles-mediated damage to cellular organelles using endoplasmic reticulum stress profile.
abstract: Gene therapy has emerged as an influential tool for treating the genetic and specific acquired disorders. Among all kinds of gene delivery systems, the cationic polymer polyethyleneimine (PEI) is considered as a promising non-viral gene delivery vector, although there are still concerns about its magnitude of cytotoxicity. While any cell insult leads to unfolded/misfolded protein accumulation and its consequent unfold protein response, evaluating the expression profile of ER-stress genes would be a sensitive indicator of cell stress. Beside cytotoxicity assays, real-time RT-PCR was used to investigate the effects of PEI nanoparticles on the endoplasmic reticulum. Treating Neuro2A cells revealed that PEI can induce cell toxicity in a concentration-dependent manner. Also, It increased the transcript levels of Grp78 (Bip), Atf4 and Chop, and splicing of Xbp1. To further optimize the transfection properties in Neuro2A cells, PEI was used to deliver a plasmid DNA containing GFP reporter. While different PEI/plasmid ratios revealed similar transfection efficiency, increasing the PEI/plasmid ratio led to induction of ER-stress markers. These results underscored that beside the effectiveness of PEI, using the lowest possible ratio of PEI/plasmid would minimize the detrimental effects of PEI on cells and confer it a beneficial therapeutic importance in nucleic acid delivery.
authors.affiliation: a Division of Pharmacology, Department of Basic Sciences, Faculty of Veterinary Medicine , Ferdowsi University of Mashhad , Mashhad , Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Dabbaghi
authors.affiliation: b Neurogenic Inflammation Research Center , Mashhad University of Medical Sciences , Mashhad , Iran.|c Department of Medical Biotechnology, School of Medicine , Mashhad University of Medical Sciences , Mashhad , Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Kazemi Oskuee
authors.affiliation: d Division of Biotechnology, Faculty of Veterinary Medicine , Ferdowsi University of Mashhad , Mashhad , Iran.
authors.forename: Khadijeh
authors.initials: K
authors.lastname: Hashemi
authors.affiliation: a Division of Pharmacology, Department of Basic Sciences, Faculty of Veterinary Medicine , Ferdowsi University of Mashhad , Mashhad , Iran.|e Biotechnology Research Group, Institute of Biotechnology , Ferdowsi University of Mashhad , Mashhad , Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Afkhami Goli
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry / Q000633:toxicity; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D006360:Heat-Shock Proteins / Q000378:metabolism; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D015388:Organelles / Q000187:drug effects* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity*; D014162:Transfection; D000071717:X-Box Binding Protein 1 / Q000378:metabolism
chemical_list: D004337:Drug Carriers; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D000071717:X-Box Binding Protein 1; C000605543:XBP1 protein, human; D011094:Polyethyleneimine; D004247:DNA
keywords: Cytotoxicity; ER-stress; polyethyleneimine; unfold protein response