title: Cellular stress induced by resazurin leads to autophagy and cell death via production of reactive oxygen species and mitochondrial impairment.
abstract: Mitochondrial bioenergetics and reactive oxygen species (ROS) often play important roles in cellular stress mechanisms. In this study we investigated how these factors are involved in the stress response triggered by resazurin (Alamar Blue) in cultured cancer cells. Resazurin is a redox reactive compound widely used as reporter agent in assays of cell biology (e.g. cell viability and metabolic activity) due to its colorimetric and fluorimetric properties. In order to investigate resazurin-induced stress mechanisms we employed cells affording different metabolic and regulatory phenotypes. In HL-60 and Jurkat leukemia cells resazurin caused mitochondrial disintegration, respiratory dysfunction, reduced proliferation, and cell death. These effects were preceded by a burst of ROS, especially in HL-60 cells which were also more sensitive and contained autophagic vesicles. Studies in Rho(0) cells (devoid of mitochondrial DNA) indicated that the stress response does not depend on the rates of mitochondrial respiration. The anti-proliferative effect of resazurin was confirmed in native acute myelogenous leukemia (AML) blasts. In conclusion, the data suggest that resazurin triggers cellular ROS production and thereby initiates a stress response leading to mitochondrial dysfunction, reduced proliferation, autophagy, and cell degradation. The ability of cells to tolerate this type of stress may be important in toxicity and chemoresistance.
authors.affiliation: Institute of Medicine, University of Bergen, Haukeland University Hospital, N-5021 Bergen, Norway.
authors.forename: Bjarte S
authors.initials: BS
authors.lastname: Erikstein
authors.forename: Hanne R
authors.initials: HR
authors.lastname: Hagland
authors.forename: Julie
authors.initials: J
authors.lastname: Nikolaisen
authors.forename: Mariola
authors.initials: M
authors.lastname: Kulawiec
authors.forename: Keshav K
authors.initials: KK
authors.lastname: Singh
authors.forename: Bjørn T
authors.initials: BT
authors.lastname: Gjertsen
authors.forename: Karl J
authors.initials: KJ
authors.lastname: Tronstad
mesh_terms: D001343:Autophagy / Q000187:drug effects*; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D019069:Cell Respiration / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D007202:Indicators and Reagents; D019169:Jurkat Cells; D008928:Mitochondria / Q000187:drug effects / Q000473:pathology*; D010078:Oxazines / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013312:Stress, Physiological; D014966:Xanthenes / Q000494:pharmacology*
chemical_list: D007202:Indicators and Reagents; D010078:Oxazines; D017382:Reactive Oxygen Species; D014966:Xanthenes; C005843:resazurin