Metal ions induced heat shock protein response by elevating superoxide anion level in HeLa cells transformed by HSE-SEAP reporter gene.
The aim of this work is to define the relationship between heat shock protein (HSP) and reactive oxygen species (ROS) in the cells exposed to different concentrations of metal ions, and to evaluate a new method for tracing the dynamic levels of cellular reactive oxygen species using a HSE-SEAP reporter gene. The expression of heat shock protein was measured using a secreted alkaline phosphatase (SEAP) reporter gene transformed into HeLa cell strain, the levels of superoxide anion (O(2)(-)) and hydrogen peroxide (H(2)O(2)) were determined by NBT reduction assay and DCFH staining flow cytometry (FCM), respectively. The experimental results demonstrated that the expression of heat shock protein induced by metal ions was linearly related to the cellular superoxide anion level before cytotoxic effects were observed, but not related to the cellular hydrogen peroxide level. The experimental results suggested that metal ions might induce heat shock protein by elevating cellular superoxide anion level, and thus the expression of heat shock protein indicated by the HSE-SEAP reporter gene can be an effective model for monitoring the dynamic level of superoxide anion and early metal-induced oxidative stress/cytotoxicity.
Yu
Zhanjiang
Z
Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100083, PR China.
Yang
Xiaoda
X
Wang
Kui
K
D002412:Cations; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry; D017930:Genes, Reporter; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008670:Metals / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects*; D020218:Response Elements / Q000235:genetics*; D013481:Superoxides / Q000378:metabolism*
D002412:Cations; D006360:Heat-Shock Proteins; D008670:Metals; D013481:Superoxides; D006861:Hydrogen Peroxide