운동신경세포주에서 재조합 인간 에리스로포이에틴(EPO)에 의한 돌연변이 SOD1/G93A단백질의 침전 억제 효과

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 rhEPO 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/mSOD1 cells, and MTT assays and BrdU-ELISAs revealed reduced cell growth and proliferation in the NSC-34/mSOD1 cell line. Incubation with 5 or 10 IU/ml rhEPO increased the viability and decreased the cox-2 expression in the dNSC-34/mSOD1 cells. Immunocytochemical staining with anti-SOD1 antibody revealed the presence of aggregates of mSOD1 protein in dNSC-34/mSOD1 cells. Incubation with10 IU/ml rhEPO reduced the proportion of cells containing such aggregates. Our findings suggest that the anti-oxidant and anti-inflammatory effects of rhEPO increase the survival of NSC-34/mSOD1 cells and reduce aggregation of the mutant SOD1 protein.