title: Hexavalent chromium induces energy metabolism disturbance and p53-dependent cell cycle arrest via reactive oxygen species in L-02 hepatocytes.
abstract: Hexavalent chromium [Cr(VI)] has become a non-negligible pollutant in the world. Cr(VI) exposure leads to severe damage to the liver, but the mechanisms involved in Cr(VI)-mediated toxicity in the liver are unclear. The present study aimed to explore whether Cr(VI) induces energy metabolism disturbance and cell cycle arrest in human L-02 hepatocytes. We showed that Cr(VI) inhibited state 3 respiration, respiratory control rate (RCR), and subsequently induced energy metabolism disturbance with decreased ATP production. Interestingly, cell cycle analysis by flow cytometry and protein expression analysis by western blotting revealed that low dose of Cr(VI) (4 uM) exposure induced S phase cell cycle arrest with decreased mediator of replication checkpoint 1 (Mrc1) and cyclin-dependent kinase 2 (CDK2), while higher doses of Cr(VI) (16, 32 uM) exposure resulted in G2/M phase arrest with decreased budding uninhibited by benzimidazoles-related 1 (BubR1) and cell division cycle 25 (CDC25). Mechanism study revealed that Cr(VI) decreased the activities of mitochondrial respiratory chain complex (MRCC) I and II, thus leading to ROS accumulation. Moreover, inhibiting ROS production by antioxidant N-acetyl-L-cysteine (NAC) rescued Cr(VI)-induced ATP depletion and cell cycle arrest. ROS-mediated p53 activation was found to involve in Cr(VI)-induced cell cycle arrest, and p53 inhibitor Pifithrin-α (PFT-α) rescued Cr(VI)-induced reduction of check point proteins Mrc1 and BubR1, thus inhibiting cell cycle arrest. In summary, the present study provides experimental evidence that Cr(VI) leads to energy metabolism disturbance and p53-dependent cell cycle arrest via ROS in L-02 hepatocytes.
authors.affiliation: Department of Health Toxicology, School of Public Health, Central South University, Changsha 410078, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Xiao
authors.forename: Xiaotao
authors.initials: X
authors.lastname: Feng
authors.forename: Ming
authors.initials: M
authors.lastname: Zeng
authors.forename: Lan
authors.initials: L
authors.lastname: Guan
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Hu
authors.forename: Caigao
authors.initials: C
authors.lastname: Zhong
mesh_terms: D000111:Acetylcysteine / Q000378:metabolism; D052160:Benzothiazoles / Q000494:pharmacology; D002274:Carcinogens, Environmental / Q000633:toxicity*; D059447:Cell Cycle Checkpoints; D002857:Chromium / Q000633:toxicity*; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004734:Energy Metabolism; D004785:Environmental Pollutants / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D014050:Toluene / Q000031:analogs & derivatives / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D052160:Benzothiazoles; D002274:Carcinogens, Environmental; D004785:Environmental Pollutants; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D002857:Chromium; C074702:chromium hexavalent ion; D014050:Toluene; C121565:pifithrin; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D000111:Acetylcysteine