The acute effect of lead acetate on glucocorticoid regulation of tyrosine aminotransferase in hepatoma cells.
Specific cellular sites of action of the environmental pollutant, lead, have not been completely defined. The present investigations were conducted to test the hypothesis that lead exposure perturbs glucocorticoid-mediated effects in hormonal target tissues. The cell culture model chosen for these investigations was the effects of lead on glucocorticoid-regulated tyrosine aminotransferase (TAT) specific activity in the H4-II-C3 hepatoma cells. Cells were treated with 300 nM-10 microM lead acetate for 24 or 48 h in absence or presence of the inducing agent, dexamethasone. Lead dose-dependently inhibited TAT specific activity up to 52% and 61% following 24 and 48 h lead treatments, respectively. These treatment times and concentrations of lead acetate did not significantly alter total cell numbers, [3H]thymidine incorporation or trypan blue exclusion. Glucocorticoid receptor-binding studies yielded a Kd = 8.3 nM and a Bmax = 290 fmol/mg protein in untreated cells versus a Kd = 9.2 nM and Bmax = 262 fmol/mg protein in cells exposed to 10 microM lead acetate for 48 h. Treatment with lead did not significantly perturb uptake of the inducing glucocorticoids or initial cytosolic receptor-binding events. To sustain induced levels of TAT, glucocorticoid must be continuously present. Following steroid withdrawal, enzyme de-induction was significantly altered in lead-treated cells. At 6 h following dexamethasone withdrawal, TAT levels had decreased to 51% of maximum in sodium acetate-treated cells. This was significantly reduced to 33% of maximum in lead acetate-treated cells. Lead treatment of HTC cells was also shown to ameliorate PMA amplification of dexamethasone-induced TAT activity. Taken together, these results suggest that acute exposure of cells to lead may inhibit processes involved in glucocorticoid-mediated enzyme induction within the hormonal target cell. Results suggest that lead may be acting to increase the turnover of TAT by actions at the transcription, translation and/or posttranslational level. Lead may also be affecting PKC-mediated phosphorylations in the glucocorticoid-TAT signal transduction system.
Heiman
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College of Pharmacy and Pharmaceutical Sciences, Florida A & M University, Tallahassee 32307, USA.
Tonner
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D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D003907:Dexamethasone / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004785:Environmental Pollutants / Q000633:toxicity; D004790:Enzyme Induction / Q000187:drug effects; D007854:Lead / Q000633:toxicity*; D008113:Liver Neoplasms / Q000201:enzymology* / Q000473:pathology; D009942:Organometallic Compounds / Q000633:toxicity*; D010766:Phosphorylation; D014176:Protein Biosynthesis / Q000187:drug effects / Q000235:genetics; D011493:Protein Kinase C / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics; D014316:Tritium / Q000378:metabolism; D014343:Trypan Blue / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014444:Tyrosine Transaminase / Q000235:genetics / Q000378:metabolism*
D004785:Environmental Pollutants; D009942:Organometallic Compounds; D011965:Receptors, Glucocorticoid; D014316:Tritium; D007854:Lead; D003907:Dexamethasone; D014444:Tyrosine Transaminase; D011493:Protein Kinase C; D014343:Trypan Blue; D013755:Tetradecanoylphorbol Acetate; C008261:lead acetate; D013936:Thymidine