title: Identification of oxidative stress and responsive genes of HepG2 cells exposed to quinocetone, and compared with its metabolites.
abstract: Quinocetone, a new quinoxaline 1,4-dioxide derivative used in food-producing animals in China, exerts genotoxic effects on HepG2 cells. It triggers significant cytotoxicity and genotoxicity in vitro, but the detailed mechanism by which quinocetone induces adverse biological effects is not yet known. We analyzed the mechanisms behind quinocetone intoxication by investigating oxidative stress based on non-enzymatic and enzymatic antioxidant activities, and by identifying differentially regulated genes of HepG2 cells exposed to quinocetone using polymerase chain reaction (PCR)-based suppression subtractive hybridization to illustrate the toxicity mechanism of quinocetone. Meanwhile, the characteristics of oxidative stress and differentially regulated genes induced by quinocetone metabolites, 1,4-bisdesoxyquinocetone and 3-methylquinoxaline-2-carboxylic acid, were investigated too. Results showed that quinocetone damaged the antioxidant defense abilities of HepG2 cells by reducing the activities of endogenous antioxidant enzymes, lowering glutathione concentration, and elevating malondialdehyde level. We identified 160 quinocetone-responsive genes that were associated with cell proliferation, glucose metabolism, oxidative stress, and apoptosis, such as NAD(P)H dehydrogenase, quinone 1; and prolyl 4-hydroxylase, beta polypeptide. The expressions of some differentially regulated genes were confirmed by real-time reverse transcription-polymerase chain reaction. However, quinocetone metabolites showed little effects on HepG2 cells. These results showed that reactive oxygen species were the key mediators of quinocetone cytotoxicity in HepG2 cells and that c-MYC-dependent activation of the mitochondrial apoptotic pathway may be associated with quinocetone-induced toxicity.
authors.affiliation: Key Laboratory of Veterinary Drug Safety Evaluation and Residues Research, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 518 Ziyue Road, Shanghai, 200241, China, z_cole@sina.com.
authors.forename: Keyu
authors.initials: K
authors.lastname: Zhang
authors.forename: Wenli
authors.initials: W
authors.lastname: Zheng
authors.forename: Haihong
authors.initials: H
authors.lastname: Zheng
authors.forename: Chunmei
authors.initials: C
authors.lastname: Wang
authors.forename: Mi
authors.initials: M
authors.lastname: Wang
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Wang
authors.forename: Lifang
authors.initials: L
authors.lastname: Zhang
authors.forename: Sui
authors.initials: S
authors.lastname: Xiao
authors.forename: Chenzhong
authors.initials: C
authors.lastname: Fei
authors.forename: Feiqun
authors.initials: F
authors.lastname: Xue
mesh_terms: D000900:Anti-Bacterial Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D004249:DNA Damage; D038181:FMN Reductase / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D005947:Glucose / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008315:Malondialdehyde / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D016877:Oxidants / Q000633:toxicity*; D018384:Oxidative Stress; D064828:Prolyl Hydroxylases / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011810:Quinoxalines / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000819:agonists / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism
chemical_list: C581286:1,4-bisdesoxyquinocetone; C581287:3-methylquinoxaline-2-carboxylic acid; D000900:Anti-Bacterial Agents; C489427:MYC protein, human; D016877:Oxidants; D016271:Proto-Oncogene Proteins c-myc; D011810:Quinoxalines; D017382:Reactive Oxygen Species; C502851:quinocetone; D008315:Malondialdehyde; D002374:Catalase; D005979:Glutathione Peroxidase; D064828:Prolyl Hydroxylases; D013482:Superoxide Dismutase; D038181:FMN Reductase; D005978:Glutathione; D005947:Glucose