| Autor: |
Mun YC; Department of Hematology and Oncology, Ewha Womans University College of Medicine, Seoul 07985, Korea., Ahn JY; Department of Hematology and Oncology, Ewha Womans University College of Medicine, Seoul 07985, Korea., Yoo ES; Department of Pediatrics, Ewha Womans University College of Medicine, Seoul 07985, Korea., Lee KE; Department of Hematology and Oncology, Ewha Womans University College of Medicine, Seoul 07985, Korea., Nam EM; Department of Hematology and Oncology, Ewha Womans University College of Medicine, Seoul 07985, Korea., Huh J; Department of Laboratory Medicine, Ewha Womans University College of Medicine, Seoul 07985, Korea., Woo HA; Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul 03760, Korea., Rhee SG; Yonsei Biomedical Research Institute, Yonsei University College of Medicine, Seoul 03722, Korea., Seong CM; Department of Hematology and Oncology, Ewha Womans University College of Medicine, Seoul 07985, Korea. |
| Abstrakt: |
NB4 cell, the human acute promyelocytic leukemia (APL) cell line, was treated with various concentrations of arsenic trioxide (ATO) to induce apoptosis, measured by staining with 7-amino-actinomycin D (7-AAD) by flow cytometry. 2', 7'-dichlorodihydro-fluorescein-diacetate (DCF-DA) and MitoSOX TM Red mitochondrial superoxide indicator were used to detect intracellular and mitochondrial reactive oxygen species (ROS). The steady-state level of SO 2 (Cysteine sulfinic acid, Cys-SO 2 H) form for peroxiredoxin 3 (PRX3) was measured by a western blot. To evaluate the effect of sulfiredoxin 1 depletion, NB4 cells were transfected with small interfering RNA and analyzed for their influence on ROS, redox enzymes, and apoptosis. The mitochondrial ROS of NB4 cells significantly increased after ATO treatment. NB4 cell apoptosis after ATO treatment increased in a time-dependent manner. Increased SO 2 form and dimeric PRX3 were observed as a hyperoxidation reaction in NB4 cells post-ATO treatment, in concordance with mitochondrial ROS accumulation. Sulfiredoxin 1 expression is downregulated by small interfering RNA transfection, which potentiated mitochondrial ROS generation and cell growth arrest in ATO-treated NB4 cells. Our results indicate that ATO-induced ROS generation in APL cell mitochondria is attributable to PRX3 hyperoxidation as well as dimerized PRX3 accumulation, subsequently triggering apoptosis. The downregulation of sulfiredoxin 1 could amplify apoptosis in ATO-treated APL cells. |
