Regulation of glutathione S-transferase enzymes in primary cultures of rat hepatocytes maintained under various matrix configurations.
Primary rat hepatocytes were cultured under various matrix and media conditions and examined after 1 week for the expression and regulation of cytosolic glutathione S-transferase (GST) enzymes. Striking effects on cell morphology were observed in relation to the different matrix conditions, whereas media effects were less prominent. Hepatocytes cultured in serum-free Dulbecco's modified Eagle's medium (DMEM) or modified Chee's medium (MCM) maintained similar levels of total GST protein regardless of the matrix configuration or corresponding cell integrity. However, HPLC analysis showed a differential expression pattern of individual GST subunits in both a time- and medium-dependent fashion. A variable, but pronounced, matrix and medium effect was observed on the induction of total GST expression by various prototypical inducers. Dexamethasone (10 microM) induced subunits A2, M1 and M2 in a medium- and matrix-dependent fashion, whereas phenobarbital (100 microM) induced significantly only subunit A2. beta-Naphthoflavone (50 microM) suppressed all GST subunit expression except subunit P1, which was induced in a matrix- and medium-dependent fashion. These studies show that total basal level expression of GSTs in vitro is reflective of a concomitant increase in mu and pi class subunits and a decrease in alpha class subunits. Moreover, the matrix and medium conditions influence both the basal and inducible expression of GST subunits in cultured rat hepatocytes.
LeCluyse
E L
EL
Department of Pharmacology, Toxicology, and Therapeutics, Center for Environmental and Occupational Health, Kansas University Medical Center, Kansas City, Kansas, USA.
Ahlgren-Beckendorf
J A
JA
Carroll
K
K
Parkinson
A
A
Johnson
J
J
D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D003907:Dexamethasone / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005110:Extracellular Space / Q000187:drug effects / Q000201:enzymology; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D019324:beta-Naphthoflavone / Q000494:pharmacology
D004791:Enzyme Inhibitors; D019324:beta-Naphthoflavone; D003907:Dexamethasone; D005982:Glutathione Transferase; D010634:Phenobarbital