title: Differential profiles of copper-induced ROS generation in human neuroblastoma and astrocytoma cells.
abstract: To determine neuronal and glial responses to copper (Cu) elevation in the CNS, human neuroblastoma and astrocytoma cells were used to compare their responses to Cu in terms of reactive oxygen species (ROS) generation and expression of enzymes responsible for anti-oxidation. Astrocytoma cells, not neuroblastoma cells, were responsive to Cu and Cu elevation was associated with ROS generation. Intracellular Cu levels as determined by inductively coupled plasma-mass spectrometry (ICP-MS), and expression levels of copper-transporting ATPase (ATP7A) and human copper transporter 1 (hCtr1) as detected by quantitative reverse transcription-polymerase chain reaction (RT-PCR), were comparable in both cell lines. Differences in Cu-induced ROS between two cell lines paralleled superoxide dismutase (SOD)-catalase expression as detected by Western blot analysis. Copper,zinc-SOD (Cu,Zn-SOD) and catalase protein levels were upregulated by Cu in neuroblastoma cells while Cu,Zn-SOD was down-regulated by Cu and catalase level was not changed in astrocytoma cells. Manganese-SOD (Mn-SOD) was not responsive to Cu in either cell line. Furthermore, 78-kDa glucose-regulated protein aggregation and upregulation were observed in Cu-treated astrocytoma cells, but not neuroblastoma cells. These data suggest that neurons use the SOD-catalase system to scavenge Cu-induced ROS while glia rely on the endoplasmic reticulum stress response to compensate for the reduction of ROS scavenging capacity.
authors.affiliation: Department of Veterinary Integrative Biosciences, Texas A and M University, College Station, TX 77843, USA. yqian@cvm.tamu.edu
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Qian
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.forename: Liz
authors.initials: L
authors.lastname: Abraham
authors.forename: Kenneth S
authors.initials: KS
authors.lastname: Ramos
authors.forename: Evelyn
authors.initials: E
authors.lastname: Tiffany-Castiglioni
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D001254:Astrocytoma / Q000473:pathology*; D015153:Blotting, Western / Q000379:methods; D002374:Catalase / Q000378:metabolism; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D003300:Copper / Q000378:metabolism; D019327:Copper Sulfate / Q000494:pharmacology*; D000079922:Copper Transport Proteins; D000073840:Copper-Transporting ATPases; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D000091342:Endoplasmic Reticulum Chaperone BiP; D005786:Gene Expression Regulation / Q000187:drug effects*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D056511:Metallochaperones; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D009447:Neuroblastoma / Q000473:pathology*; D012333:RNA, Messenger / Q000096:biosynthesis; D017382:Reactive Oxygen Species / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: C474500:ATOX1 protein, human; D027682:Cation Transport Proteins; D000079922:Copper Transport Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D056511:Metallochaperones; D018832:Molecular Chaperones; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D003300:Copper; D002374:Catalase; D013482:Superoxide Dismutase; D000251:Adenosine Triphosphatases; C081184:ATP7A protein, human; D000073840:Copper-Transporting ATPases; D019327:Copper Sulfate