title: Recombinant human erythropoietin reduces aggregation of mutant Cu/Zn-binding superoxide dismutase (SOD1) in NSC-34 cells.
abstract: Human erythropoietin (hEPO) has multiple actions in non-hematopoietic tissues, including neurotrophic, anti-oxidant, anti-apoptotic, and anti-inflammatory effects. To examine the effect of EPO in an vitro model of amyotrophic lateral sclerosis (ALS), we stably overexpressed wild SOD1 and a mutant form, SOD1/G93A, in NSC-34 motoneuron-like cells. Transformants harboring the wild and mutant forms of SOD1 were selected by G418 selection and immunoblot analysis. RT-PCR analysis showed that cox-2 expression was increased in the NSC-34/mSOD1s, and MTT assays and BrdU-ELISAs revealed reduced cell growth and proliferation in the NSC-34/mSOD1 cell line. Incubation with 5 or 10IU/mL rhEPO increased the viability and decreased the cox-2 expression in the dNSC-34/mSOD1s cells. Immunocytochemical staining with anti-SOD1 antibody revealed the presence of aggregates of mSOD1 protein in dNSC-34/mSOD1 cells. Incubation with10IU/mL rhEPO reduced the proportion of cells containing such aggregates. Our findings suggest that the anti-oxidant and anti-inflammatory effects of EPO increase the survival of NSC-34/mSOD1 cells and reduce aggregation of the mutant SOD1 protein.
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, Seoul, Republic of Korea; Department of Biology, College of Natural Sciences, Chosun University, Gwangju, 501-759, Republic of Korea.
authors.forename: Goang-Won
authors.initials: GW
authors.lastname: Cho
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, Seoul, Republic of Korea.
authors.forename: Ga-Young
authors.initials: GY
authors.lastname: Kim
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, Seoul, Republic of Korea.
authors.forename: Soojeong
authors.initials: S
authors.lastname: Baek
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, Seoul, Republic of Korea.
authors.forename: Heejaung
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Physical Medicine & Rehabilitation, College of Medicine, Hanyang University, Seoul, Republic of Korea.
authors.forename: Taikon
authors.initials: T
authors.lastname: Kim
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, Seoul, Republic of Korea.
authors.forename: Hee Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, Seoul, Republic of Korea. Electronic address: kimsh1@hanyang.ac.kr.
authors.forename: Seung Hyun
authors.initials: SH
authors.lastname: Kim
mesh_terms: D000818:Animals; D000963:Antimetabolites; D001973:Bromodeoxyuridine; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000096:biosynthesis; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004921:Erythropoietin / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism*; D016296:Mutagenesis; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000963:Antimetabolites; D018076:DNA, Complementary; D009841:Oligonucleotides; D011994:Recombinant Proteins; D004921:Erythropoietin; D012313:RNA; D051546:Cyclooxygenase 2; D013482:Superoxide Dismutase; D002795:Choline O-Acetyltransferase; D001973:Bromodeoxyuridine