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Selected Publication:
Steiner, E; Kleinhappl, B; Gutschi, A; Marth, E.
Analysis of hsp70 mRNA levels in HepG2 cells exposed to various metals differing in toxicity
Toxicol Lett. 1998; 96-97(2):169-176
Doi: 10.1016%2FS0378-4274%2898%2900065-4
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- Co-authors Med Uni Graz
- Kleinhappl Barbara
- Marth Egon
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- Abstract:
- Human hepatoma cells (HepG2) were exposed to several heavy metal salts and the induction of heat shock protein 70 (hsp70) mRNA was analysed by the reverse transcriptase-polymerase chain reaction (RT-PCR). Metals were added to the cell medium at concentrations ranging from 0.1 to 100 microM and incubation was continued for 4 h. In addition we analysed the time dependence of hsp70 induction by adding each metal at a certain concentration followed by an incubation for 0.5 to 24 h. CdCl2, NaAsO2, AgNO3 could be classified as very strong inducers (20-, 13- and 10-fold above control level) and they reached their maximum level of induction at 1-10 microM after 2 h. CuCl2, MnCl2, Pb(NO3)2, TlNO3, CoCl2 and NiCl2 were also strong inducing agents, giving a 4-6 fold induction at 10-100 microM after 4-8 h. ZnSO4, Hg(NO3)2 and AlCl3 were only weak inducers (1.5-2 fold at 50-100 microM after 4-8 h) of hsp70 mRNA. Cytotoxic effects (measured by release of lactate dehydrogenase) could only be detected for 100 microM Hg2+ after 4 h and when the cells were incubated with 5 microM Cd2+ for more than 8 h. We also tested a few combinations of these heavy metal salts for their hsp70-inducing ability. Zn2+ and Mn2+ were able to diminish Cd2+ induced hsp70 mRNA levels by 65%. Ag+ mediated induction was reduced by 40% when combined with Cu2+, whereas Hg2+ increased induction by Ag+ about 3-fold and led to a dramatic decrease in cell viability. In our study we were able to demonstrate that the analysis of hsp70 mRNA levels in chemically stressed HepG2 cells by RT-PCR can be a valuable tool for studying mechanisms of toxicity associated with elevated expression of hsp70.
- Find related publications in this database (using NLM MeSH Indexing)
- Cadmium Chloride - toxicity
- Carcinoma, Hepatocellular - metabolism
- Gene Expression - drug effects
- HSP70 Heat-Shock Proteins - biosynthesis
- Humans - biosynthesis
- Liver Neoplasms - metabolism
- Metals, Heavy - toxicity
- RNA, Messenger - metabolism
- Reverse Transcriptase Polymerase Chain Reaction - metabolism
- Tumor Cells, Cultured - metabolism
- Find related publications in this database (Keywords)
- Heavy Metals
- Hsp70
- RT-PCR
- HepG2