Cloning of the 11beta-hydroxysteroid dehydrogenase (11beta-HSD)-2 gene in the baboon: effects of estradiol on promoter activity of 11beta-HSD-1 and -2 in placental JEG-3 cells

Biochim Biophys Acta. 1999 Jan 18;1444(1):101-10. doi: 10.1016/s0167-4781(98)00248-6.


In the baboon, estrogen regulated 11beta-hydroxysteroid dehydrogenase (11beta-HSD) catalyzed metabolism of cortisol and cortisone by the placenta is an important component in the sequence of events regulating the fetal pituitary-adrenocortical axis. The present study was designed to isolate and sequence the promoter region of the baboon 11beta-HSD-2 gene and to produce constructs of this gene and the 1.7 kb fragment of 5'-flanking region of baboon 11beta-HSD-1 isolated previously in order to determine whether the promoters of these two genes were activated in human placental JEG-3 cells and whether expression could be modulated by estradiol. The 11beta-HSD-2 genomic DNA was isolated from a baboon kidney genomic library using a human 11beta-HSD-2 cDNA as a probe. The sequence of a 1.2 kb fragment of the 5'-flanking region showed extensive homology with that published by others for human 11beta-HSD-2, particularly in exon 1 (>95%) and in the proximal promoter (>90%). Primer extension confirmed that the baboon 11beta-HSD-2 gene has multiple transcriptional start sites which are preceded by a GC box. To determine promoter activity of 11beta-HSD-2 and -1, the 5'-flanking regions of these genes were subcloned into luciferase reporter pGL3 vectors, transiently transfected into human placental JEG-3 cells, and then incubated for 16-18 h in the presence or absence of 10-8 M 17beta-estradiol or 17alpha-estradiol. To augment the low level of estrogen receptor (ER) in JEG cells, promoter activity studies were also performed in JEG cells co-transfected with an expression vector containing the human ER cDNA. The promoters of both 11beta-HSD-1 and -2 were activated following transient transfection into JEG-3 cells although basal activity of 11beta-HSD-2 (87+/-21 RLU/microg protein) always exceeded (P<0.05) that of 11beta-HSD-1 (37+/-7). In the absence of co-transfected ER, basal promoter activities of both 11beta-HSD genes were not altered by 17beta-estradiol. In contrast, in cells co-transfected with ER, 17beta-estradiol but not 17alpha-estradiol increased (P<0.05) basal promoter activities of 11beta-HSD-1 and -2 by 8.1+/-1.5 and 8.3+/-2. 0 fold, respectively. Collectively, these findings indicate that the promoter region of the baboon 11beta-HSD-2 gene is comparable to that in the human and that the 5'-flanking region of both the baboon 11beta-HSD-1 and -2 genes were active when transiently transfected into JEG-3 cells and that activation could be enhanced by estradiol in the presence of an estrogen receptor.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 11-beta-Hydroxysteroid Dehydrogenases
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • DNA / chemistry
  • Estradiol / pharmacology*
  • Gene Expression Regulation / drug effects
  • Humans
  • Hydroxysteroid Dehydrogenases / biosynthesis
  • Hydroxysteroid Dehydrogenases / genetics*
  • Kidney / metabolism
  • Molecular Sequence Data
  • Papio
  • Placenta / metabolism
  • Promoter Regions, Genetic / drug effects*
  • RNA / isolation & purification
  • Receptors, Estrogen / biosynthesis
  • Receptors, Estrogen / genetics
  • Sequence Homology, Nucleic Acid
  • Transfection


  • Receptors, Estrogen
  • Estradiol
  • RNA
  • DNA
  • Hydroxysteroid Dehydrogenases
  • 11-beta-Hydroxysteroid Dehydrogenases