Different induction of metallothioneins and Hsp70 and presence of the membrane transporter ZnT-1 in HepG2 cells exposed to copper and zinc.
Eukaryotic cells respond to stressful environmental stimuli, such as toxic concentrations of heavy metals, by rapidly synthesising defence proteins: the metallothioneins (MT) and the heat shock protein 70 (Hsp70). In this study we have analysed how the human hepatoblastoma cell line HepG2 responds to exposure to excess copper (30 microg/ml) and zinc (50 microg/ml) for long exposure times (48 and 72 h). Accumulation of the two metals, as measured by ICP-AES, was time-dependent reaching a plateau after 72 h. HepG2 cells responded by dramatically increasing levels of MT during stress, mostly during zinc exposure. A time lag in Hsp70 induction was observed as the levels of this protein increased only after removal of the stress from culture medium (recovery) for 24 h, thus suggesting that the two defence mechanisms are not coordinated in a metal-induced stress response. Moreover in HepG2 cells, immunochemical and fluorescence techniques showed the presence and the localisation of the zinc membrane exporter ZnT-1 as a further mechanism of defence/homeostasis against zinc toxicity.
Urani
C
C
Dipartimento di Scienze dell'Ambiente e del Territorio, Università degli Studi di Milano Bicocca, p.zza della Scienza 1, 20126 Milan, Italy. chiara.urani@unimib.it
Calini
V
V
Melchioretto
P
P
Morazzoni
F
F
Canevali
C
C
Camatini
M
M
D045744:Cell Line, Tumor; D003300:Copper / Q000032:analysis / Q000633:toxicity*; D019084:Fluorescent Antibody Technique, Indirect; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis*; D018197:Hepatoblastoma; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008113:Liver Neoplasms; D008565:Membrane Proteins / Q000378:metabolism*; D008668:Metallothionein / Q000096:biosynthesis*; D013054:Spectrophotometry, Atomic / Q000379:methods; D013997:Time Factors; D015032:Zinc / Q000032:analysis / Q000633:toxicity*
D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D003300:Copper; D008668:Metallothionein; D015032:Zinc