TY - JOUR
T1 - Selective expression of cytochrome P450 isozymes by 4-n-alkyl-methylenedioxybenzenes in rat lung cells
AU - Marcus, Craig B.
AU - Wilson, Neil M.
AU - Keith, Ingegerd M.
AU - Jefcoate, Colin R.
AU - Omiecinski, Curtis J.
N1 - Funding Information:
This work was supported in part by National Institutes of Health Grants GM-32281 andES-04696 (to C.J.O.) andCA-16265 (to C.R.J.), and by American Cancer Society Institutional Grant IN-17 (to C.B.M.). N.M.W. was supported in part by National Institutes of Environmental Health Sciences Training Grant ES-07015. Work conducted by Dr. Ingegerd Keith was supported by the College of Agricultural and Life Sciences, University of Wisconsin. The authors thank Dr. Burt Olson, Karen Rudd, and Robert Kalwinsky for technical assistance in portions of this work, and Dr. Maro Christou for analyzing the DMBA HPLC assays.
PY - 1990/2/15
Y1 - 1990/2/15
N2 - Rat lung preparations were analyzed for cytochrome P450 expression as a function of pretreatment with 1 mmol/kg/day of various substituted n-alkyl-methylenedioxybenzenes (MDBs). Lung P450s were quantitated by Western blotting and visualized by immunocytochemical techniques using polyclonal antibodies directed against P450IA1/IA2 and P450IIB1/IIB2. Results demonstrated that n-hexyland n-octyl-MDB (but not n-butyl-MDB) increased P450IA1 in lung microsomes (50 pmol/mg microsomal protein) compared to untreated controls and n-butyl-MDB-treated animals (6 pmol/mg). However, treatment with all of these MDBs concomitantly decreased P450IIB1 in rat lung microsomes, from 40 pmol/mg in untreated control lungs to below the detection limit of this method (<2 pmol/mg). In vitro metabolism assays with rat lung microsomes, utilizing 7,12-dimethylbenz[a]anthracene as substrate, confirmed the increase in P450IA1 by selected MDBs and the concomitant decrease of catalytically active P450IIB1. Immunocytochemistry of rat lungs with these same IgGs indicated selective cellular responses. n-Hexyl-MDB (but not n-butyl-MDB) increased P450IA1-like immunoreactivity, but the increase was specifically localized to Clara cells. Immunocytochemical results further showed that P450IIB1-like immunoreactivity disappeared entirely from alveolar type II cells, yet appeared unchanged in Clara cells relative to untreated controls. Increases in P450IA1 due to treatment with n-hexyl-MDB were associated with increases in P450IA1 mRNA as indicated by Northern blot experiments. In contrast, the decreased levels of P450IIB1, consequent to treatment with n-alkyl-MDBs, were not associated with altered levels of pulmonary P450IIB1 mRNA as determined by Northern blotting and solution hybridization assays. These results imply that n-alkyl-MDBs regulate pulmonary cytochromes P450 by both transcriptional and post-transcriptional mechanisms.
AB - Rat lung preparations were analyzed for cytochrome P450 expression as a function of pretreatment with 1 mmol/kg/day of various substituted n-alkyl-methylenedioxybenzenes (MDBs). Lung P450s were quantitated by Western blotting and visualized by immunocytochemical techniques using polyclonal antibodies directed against P450IA1/IA2 and P450IIB1/IIB2. Results demonstrated that n-hexyland n-octyl-MDB (but not n-butyl-MDB) increased P450IA1 in lung microsomes (50 pmol/mg microsomal protein) compared to untreated controls and n-butyl-MDB-treated animals (6 pmol/mg). However, treatment with all of these MDBs concomitantly decreased P450IIB1 in rat lung microsomes, from 40 pmol/mg in untreated control lungs to below the detection limit of this method (<2 pmol/mg). In vitro metabolism assays with rat lung microsomes, utilizing 7,12-dimethylbenz[a]anthracene as substrate, confirmed the increase in P450IA1 by selected MDBs and the concomitant decrease of catalytically active P450IIB1. Immunocytochemistry of rat lungs with these same IgGs indicated selective cellular responses. n-Hexyl-MDB (but not n-butyl-MDB) increased P450IA1-like immunoreactivity, but the increase was specifically localized to Clara cells. Immunocytochemical results further showed that P450IIB1-like immunoreactivity disappeared entirely from alveolar type II cells, yet appeared unchanged in Clara cells relative to untreated controls. Increases in P450IA1 due to treatment with n-hexyl-MDB were associated with increases in P450IA1 mRNA as indicated by Northern blot experiments. In contrast, the decreased levels of P450IIB1, consequent to treatment with n-alkyl-MDBs, were not associated with altered levels of pulmonary P450IIB1 mRNA as determined by Northern blotting and solution hybridization assays. These results imply that n-alkyl-MDBs regulate pulmonary cytochromes P450 by both transcriptional and post-transcriptional mechanisms.
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U2 - 10.1016/0003-9861(90)90544-9
DO - 10.1016/0003-9861(90)90544-9
M3 - Article
C2 - 2306118
AN - SCOPUS:0025057321
SN - 0003-9861
VL - 277
SP - 17
EP - 25
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -