Heat shock protein 70 as an indicator of early lung injury caused by exposure to arsenic.
Heat shock proteins (HSPs) are a family of highly conserved proteins that are induced by a number of stresses including toxic metals. Heat shock proteins expression has been reported to be an early and sensitive biomarker of cell stress. Arsenic is a naturally occurring metal that exists widely in the environment and is used in several industries. Exposure to arsenic is associated with the development of pulmonary cancers. We monitored changes in Hsp70 and markers of oxidative injury induced by arsenic in human pulmonary epithelial cells (BEAS-2B). Hsp70 protein, mRNA and reactive oxygen species (ROS) generation were measured after exposing cells to arsenic as markers of injury. Hsp70 protein expression showed significant 7.9-fold and 31.5-fold increase using Western blotting and ELISA assay, respectively, at a 50 microM As(III) with a 12 h exposure and an 12 h recovery time. Hsp70A and Hsp70B mRNA expression showed a two-fold increase and Hsp70C mRNA expression showed a six-fold increase. As(III)-induced Hsp70 protein expression was inhibited significantly by catalase and NAC, indicating mediation of ROS in Hsp70 expression. Intracellular glutathione (GSH) was significantly depleted by As(III) exposure. Lipid peroxidation by-product, 8-isoprostane, was increased six-fold at 24 h exposure to 20 microM As(III). Electron spin resonance and confocal microscope studies also showed As(III)-stimulated ROS generation. These results suggest that cellular injury by arsenic is mediated through ROS generation resulting in the expression of Hsp70. It is possible that Hsp70 may prove to be a sensitive biomarker for arsenic exposure with other markers of oxidative stress in human serum.
Han
Sung Gu
SG
Pathology and Physiology Research Branch, National Institute for Occupational Safety and Health, 1095 Willowdale Road, Morgantown, West Virginia 26505, USA.
Castranova
Vince
V
Vallyathan
Val
V
D000975:Antioxidants / Q000494:pharmacology; D001151:Arsenic / Q000008:administration & dosage / Q000633:toxicity*; D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D018076:DNA, Complementary / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D017665:Hydroxyl Radical / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008168:Lung / Q000187:drug effects* / Q000378:metabolism*; D055370:Lung Injury; D018384:Oxidative Stress / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D026361:Reactive Nitrogen Species / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D000975:Antioxidants; D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D017665:Hydroxyl Radical; D005978:Glutathione; D001151:Arsenic