title: Regulation of aromatase in estrogen-producing cells.
abstract: Human adipose stromal cells in monolayer culture aromatize androstenedione to estrone. The rate of aromatization is stimulated 20- to 30-fold by glucocorticoids when fetal calf serum is present in the culture medium and by dibutyryl cyclic AMP in the absence of serum. The action of dibutyryl cyclic AMP to stimulate aromatase activity is potentiated markedly by phorbol esters and inhibited by growth factors, such as EGF. In order to investigate the mechanisms underlying this multifactorial regulation, we have prepared polyclonal and monoclonal antibodies specific for aromatase cytochrome P-450. By use of these antibodies it was demonstrated that the action of these various factors to regulate aromatase activity was caused by alterations in the rate of synthesis of aromatase cytochrome P-450, whereas the synthesis of the reductase component of the aromatase enzyme complex was relatively unaffected. The changes in the rate of synthesis of aromatase cytochrome P-450 were, in turn, reflective of changes in the levels of translatable mRNA specific for this protein. In order to analyze the levels of aromatase cytochrome P-450 mRNA directly, we have isolated a cloned cDNA insert complementary to the mRNA encoding aromatase cytochrome P-450, by screening a lambda gt 11 human placental cDNA library utilizing the polyclonal anti-aromatase P-450 IgG. Use of this cDNA probe in Northern analysis of RNA extracted from human adipose stromal cells revealed that the changes in translatable mRNA resulting from incubation of the cells with the various regulatory factors were due to changes in the absolute levels of mRNA encoding this protein.
authors.affiliation: Cecil H. and Ida Green Center for Reproductive Biology Sciences, Department of Obstetrics and Gynecology, University of Texas Health Science Center, Dallas 75235-9051.
authors.forename: C R
authors.initials: CR
authors.lastname: Mendelson
authors.forename: C T
authors.initials: CT
authors.lastname: Evans
authors.forename: E R
authors.initials: ER
authors.lastname: Simpson
mesh_terms: D000273:Adipose Tissue / Q000201:enzymology*; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004274:DNA, Recombinant; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004967:Estrogens / Q000096:biosynthesis*; D005260:Female; D006801:Humans; D007163:Immunosorbent Techniques; D009693:Nucleic Acid Hybridization; D010703:Phorbol Esters / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D004274:DNA, Recombinant; D004967:Estrogens; D010703:Phorbol Esters; D012333:RNA, Messenger; D004815:Epidermal Growth Factor; D003994:Bucladesine; D004247:DNA; D001141:Aromatase