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Abstract

Exposure of Hep3B cells to metalloporphyrins (tinprotoporphyrin and heme) or cobalt chloride resulted in the production of a significant number of heme oxygenase transcripts, erythropoietin transcripts or both, as indicated by in situ hybridization. Exposure to heme 10 μmol/L resulted in a 30-fold to 40-fold increase in cells expressing erythropoietin messenger RNA (erythropoietin-positive cells) by 6 hr; this increased level remained elevated for 24 hr. Tin-protoporphyrin (10 μmol/L) produced an eightfold to 10-fold increase in erythropoietin RNA within 40 min. This value then returned to control levels by 60 min. Exposure to cobalt chloride (100 μmol/L) resulted in a 20-fold to 30-fold increase in erythropoietin expression for 5 to 20 min, returning to control by 40 min. Additionally, nuclear runoff assays demonstrated that the increase in heme oxygenase or erythropoietin messenger RNA accumulation by cobalt chloride appeared to be a result of stimulated transcription of the heme oxygenase and erythropoietin genes. However, the pattern for heme oxygenase messenger RNA induction was different from that for erythropoietin expression. Heme produced an immediate expression of heme oxygenase RNA (50-fold within 5 min) and a second sustained response during the next 24 hr. Tin-protoporphyrin also produced an immediate response (40-fold within 5 min) and remained elevated (20-fold) for 6 hr. Cobalt chloride produced a 22-fold increase within 20 min and returned to the control value by 1 hr. Thus both erythropoietin and heme oxygenase genes appear to be expressed after treatment with tin-protoporphyrin, heme or cobalt chloride; however, the time and patterns of expression are different. The most significant feature of coexpression appears to be that induced by heme, where heme oxygenase is immediately expressed (5 min) with a second sustained expression during the next 10 to 24 hr, whereas erythropoietin sustained expression for 3 to 24 hr. (HEPATOLOGY 1993;17:861–868.)