Copper toxicity affects proliferation and viability of human hepatoma cells (HepG2 line).
In Wilson's disease and Indian childhood cirrhosis (ICC) copper accumulates in the liver resulting in poor hepatocyte regeneration and fibrosis. An inhibition of hepatocyte proliferation and an increase in cell death could account for these outcomes. To establish how the toxicity of this metal ion impacts upon the proliferation and viability of the HepG2 cells they were cultured in 4-32 microM copper(II) sulphate (CuSO4)). These levels were comparable to the circulatory and tissue concentrations of copper recorded for these two diseases. Specific uptake comparable to levels of copper recorded in the livers of patients with Wilson's disease and ICC was measured in the HepG2 cells. After 48 h acid vesicle function increased from 4 to 32 microM Cu2+ but significantly declined at 64 microM compared to the controls. Lysosomal acid phosphatase showed a concentration dependent decline in activity at 72 h. Cellls exposed to 64 microM Cu2+ had a potential doubling time (Tpot) 21 h longer than the control cells due to a prolonged DNA synthesis phase. At 64 microM Cu2+, increases of necrosis up to 18% were seen whereas comparable levels of apoptotic and necrotic cells (<5%) were seen below this concentration. Chronic exposure over 8 weeks impaired colony-forming efficiency at concentrations of 16 microM Cu2+ and above. This study suggests that when liver cells sequester large amounts of copper, the toxic effects include delayed cell-cycle progression, a gradual loss of replicative capacity, and an increased incidence of cell death.
Aston
N S
NS
University Division of Child Health, Stephenson Unit, The Children's Hospital, Sheffield, S10 2TH, UK.
Watt
N
N
Morton
I E
IE
Tanner
M S
MS
Evans
G S
GS
D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D002648:Child; D002675:Child, Preschool; D003300:Copper / Q000493:pharmacokinetics / Q000633:toxicity*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D016195:G2 Phase / Q000187:drug effects; D006527:Hepatolenticular Degeneration / Q000097:blood; D006801:Humans; D007120:Immunochemistry; D008103:Liver Cirrhosis / Q000097:blood; D008113:Liver Neoplasms / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects; D009336:Necrosis; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000473:pathology; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D014410:Tumor Stem Cell Assay
D003300:Copper; D016650:Fluorescein-5-isothiocyanate