Sulfotransferase gene expression in primary cultures of rat hepatocytes

• Published in: Biochemical pharmacology 1996-11, Vol.52 (10), p.1621-1630
• Main Authors: Liu, Lan, LeCluyse, Edward L., Liu, Jie, Klaassen, Curtis D.
• Format: Article
• Language: English
• Subjects: Albumins - genetics; Analytical, structural and metabolic biochemistry; Animals; Arylsulfotransferase - genetics; Base Sequence; Biological and medical sciences; Cells, Cultured; conditioned culture; dexamethasone; DNA Probes - genetics; Enzymes and enzyme inhibitors; Female; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic; Liver - cytology; Liver - enzymology; Liver - metabolism; Liver. Bile. Biliary tracts; Male; matrigel; phenol (ST1A1) mRNA; rat hepatocyte primary culture; Rats; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sex Characteristics; Sulfotransferase mRNAs; Sulfotransferases - genetics; Transferases; Vertebrates: digestive system
• ISSN: 0006-2952; 1873-2968
• DOI: 10.1016/S0006-2952(96)00569-2

Hepatocyte cultures have been used in pharmacotoxicological studies, and sulfotransferases (ST) are important drug-metabolizing enzymes in liver. The expression of sulfotransferases in hepatocyte cultures has not been examined systematically. In the present study, the mRNA levels of different sulfotransferases in male and female rat hepatocytes were examined by northern-blot analyses. Various culture conditions such as different matrices (collagen, matrigel, collagen sandwich, or co-culture with epithelial cells), medium (Waymouth's MB 752/1 and Modified Chee's Medium) and glucocorticoid supplementation (dexamethasone, 0.1 μM) were compared. Phenol ST (ST1A1) mRNA levels decreased to about 50% of initial mRNA levels within 10 hr of culture. At 96 hr, ST1 Al mRNA levels were approximately 20% of initial values when cultured on collagen, matrigel or co-culture. The two media did not differ in ability to maintain ST1A1 mRNA levels in the absence of dexamethasone (DEX); however, DEX addition to either medium resulted in ST1A1 mRNA levels greater than 100% of the initial mRNA levels at 96 hr, with the greatest increase observed using the matrigel substratum and Chee's medium. In the absence of DEX, the mRNA levels of N-hydroxy-2acetylaminofluorene Sulfotransferase (ST1C1), estrogen sulfotransferase (ST1E2) and hydroxysteroid sulfotransferase (ST-20/21, ST-40/41, ST-60) fell to approximately 20% of their initial levels within 24 hr, and to less than 5% at 96 hr. The loss of expression of these sulfotransferases was observed with all culture conditions. Addition of DEX to the media resulted in ST-40/41 and ST-60 mRNA expression at 20 and 35% of their initial values, respectively, in cultures maintained on matrigel and Chee's medium at 96 hr. These data suggest that sulfotransferases lose their constitutive expression in hepatocyte culture, but retain their inducibility.