Regulation of cytochrome P-450c by glucocorticoids and polycyclic aromatic hydrocarbons in cultured fetal rat hepatocytes

James Michael Mathis, Russell A. Prough, Ronald N. Hines, Edward Bresnick, Evan R. Simpson

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Abstract

The actions of polycyclic aromatic hydrocarbons and glucocorticoids to regulate the synthesis of cytochrome P-450c (the major isozyme induced by polycyclic aromatic hydrocarbons) were investigated in fetal rat hepatocytes maintained in primary monolayer culture. Treatment of hepatocytes in culture with 1,2-benzanthracene resulted in a 50-fold increase in 7-ethoxycoumarin O-deethylase activity. The level of P-450c increased in the cells in a time-dependent fashion as determined by immunoelectrophoretic analysis. The inductive effect of BA was potentiated approximately 1.6- to 2.3-fold when 1 μm dexamethasone was included in the culture medium. However, dexamethasone alone had little or no effect on the induction of P-450c. The rate of synthesis of P-450c was examined by immunoisolation of the specific isozyme from total cellular proteins radiolabeled with [35S]methionine and from the protein products formed during in vitro translation of the isolated mRNA. In addition, the amount of mRNA specific for cytochrome P-450c was determined by Northern blot analysis of RNA extracted from cultured cells. The changes in the rates of synthesis and mRNA levels were found to parallel the changes in enzyme activity. The concentration of dexamethasone required to cause a half-maximal increase in P-450c content in the presence of 1,2-benzanthracene was between 10-8 and 10-7 m. It is concluded that glucocorticoids act synergistically with polycyclic aromatic hydrocarbons to increase the levels of P-450c expressed in the fetal rat liver, and that this action is likely mediated by the classical type II glucocorticoid receptor.

Original languageEnglish
Pages (from-to)439-448
Number of pages10
JournalArchives of Biochemistry and Biophysics
Volume246
Issue number1
DOIs
StatePublished - 1 Jan 1986

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Polycyclic Aromatic Hydrocarbons
Cytochromes
Dexamethasone
Glucocorticoids
Rats
Hepatocytes
Messenger RNA
Isoenzymes
7-Alkoxycoumarin O-Dealkylase
Glucocorticoid Receptors
Enzyme activity
Protein Biosynthesis
Northern Blotting
Methionine
Liver
Culture Media
Cultured Cells
Monolayers
Proteins
Cells

Cite this

Mathis, James Michael ; Prough, Russell A. ; Hines, Ronald N. ; Bresnick, Edward ; Simpson, Evan R. / Regulation of cytochrome P-450c by glucocorticoids and polycyclic aromatic hydrocarbons in cultured fetal rat hepatocytes. In: Archives of Biochemistry and Biophysics. 1986 ; Vol. 246, No. 1. pp. 439-448.
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Regulation of cytochrome P-450c by glucocorticoids and polycyclic aromatic hydrocarbons in cultured fetal rat hepatocytes. / Mathis, James Michael; Prough, Russell A.; Hines, Ronald N.; Bresnick, Edward; Simpson, Evan R.

In: Archives of Biochemistry and Biophysics, Vol. 246, No. 1, 01.01.1986, p. 439-448.

Research output: Contribution to journalArticle

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