title: Development of HSPA1A promoter-driven luciferase reporter gene assays in human cells for assessing the oxidative damage induced by silver nanoparticles.
abstract: The exponential increase in the total number of engineered nanoparticles in consumer products requires novel tools for rapid and cost-effective toxicology screening. In order to assess the oxidative damage induced by nanoparticles, toxicity test systems based on a human HSPA1A promoter-driven luciferase reporter in HepG2, LO2, A549, and HBE cells were established. After treated with heat shock and a group of silver nanoparticles (AgNPs) with different primary particle sizes, the cell viability, oxidative damage, and luciferase activity were determined. The time-dependent Ag(+) ions release from AgNPs in cell medium was also evaluated. Our results showed that heat shock produced a strong time-dependent induction of relative luciferase activity in the four luciferase reporter cells. Surprisingly, at 4h of recovery, the relative luciferase activity was >98× the control level in HepG2-luciferase cells. Exposure to different sizes of AgNPs resulted in activation of the HSPA1A promoter in a dose-dependent manner, even at low cytotoxic or non-cytotoxic doses. The smaller (5nm) AgNPs were more potent in luciferase induction than the larger (50 and 75nm) AgNPs. These results were generally in accordance with the oxidative damage indicated by malondialdehyde concentration, reactive oxygen species induction and glutathione depletion, and Ag(+) ions release in cell medium. Compared with the other three luciferase reporter cells, the luciferase signal in HepG2-luciferase cells is obviously more sensitive and stable. We conclude that the luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the oxidative damage induced by AgNPs.
authors.affiliation: School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu, China; Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, School of Public Health, Soochow University, Suzhou 215123, China. Electronic address: llxin@suda.edu.cn.
authors.forename: Lili
authors.initials: L
authors.lastname: Xin
authors.affiliation: Suzhou Center for Disease Prevention and Control, 72 Sanxiang Road, Suzhou, Jiangsu, China.
authors.forename: Jianshu
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Radiation Medicine and Protection & School for Radiological and Interdisciplinary Sciences (RAD-X), Soochow University, 215123, China.
authors.forename: Leshuai W
authors.initials: LW
authors.lastname: Zhang
authors.affiliation: School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu, China.
authors.forename: Bizhong
authors.initials: B
authors.lastname: Che
authors.affiliation: School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu, China.
authors.forename: Guangzhu
authors.initials: G
authors.lastname: Dong
authors.affiliation: Suzhou Industrial Park Centers for Disease Control and Prevention, 58 Suqian Road, Suzhou, Jiangsu, China.
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Fan
authors.affiliation: Suzhou Industrial Park Centers for Disease Control and Prevention, 58 Suqian Road, Suzhou, Jiangsu, China.
authors.forename: Kaiming
authors.initials: K
authors.lastname: Cheng
mesh_terms: D000072283:A549 Cells; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D017930:Genes, Reporter / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D008156:Luciferases / Q000187:drug effects; D008315:Malondialdehyde / Q000378:metabolism; D053768:Metal Nanoparticles / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects*; D010316:Particle Size; D011401:Promoter Regions, Genetic / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D012834:Silver / Q000633:toxicity*; D018675:Toxicity Tests
chemical_list: D018840:HSP70 Heat-Shock Proteins; C490786:HSPA1A protein, human; D017382:Reactive Oxygen Species; D012834:Silver; D008315:Malondialdehyde; D008156:Luciferases; D005978:Glutathione
keywords: HSPA1A; Luciferase reporter; Oxidative stress; Silver nanoparticles