Characterization of 17 beta-hydroxysteroid dehydrogenase type 1 in choriocarcinoma cells: regulation by basic fibroblast growth factor.
17 beta-Hydroxysteroid dehydrogenase type 1 (17-HSD type 1) is a steroidogenic enzyme catalyzing reversible interconversion of estradiol and estrone. 17-HSD type 1 is actively expressed in human placenta. We characterized 17-HSD type 1 expression and its regulation by basic fibroblast growth factor (bFGF) in JAR, JEG-3 and BeWo choriocarcinoma cell lines. Based on Southern and Northern analysis, as well as measurement of catalytic activity and immunoreactive protein, all the choriocarcinoma cell lines contained and expressed the gene coding for 17-HSD type 1, identical to that of normal human cells. However, the cell lines showed marked quantitative differences in the levels of expression of the enzyme, being lowest in JAR cells and highest in BeWo cells, as measured by immunofluorometric assay, Northern analysis and catalytic activity. These differences in the basal level of expression were most probably not based on any sequence differences in the putative proximal promoter area of the gene in different cell lines, since no dissimilarities were observed in the 806 bp region upstream from the transcription start site of 1.3 kb mRNA coding for 17-HSD type 1 except for frequent polymorphism characteristic of normal human cells using polymerase chain reaction/single-strand conformation polymorphism (PCR-SSCP) analysis. The reductive (estrone-->estradiol) activity was about 4-7 times higher compared with the oxidative activity (estradiol-->estrone) in all the cell lines studied, indicating that in these choriocarcinoma cell lines, 17-HSD activity favours estradiol formation.(ABSTRACT TRUNCATED AT 250 WORDS)
Lewintre
E J
EJ
Department of Clinical Chemistry, University of Oulu, Finland.
Orava
M
M
Peltoketo
H
H
Vihko
R
R
D015067:17-Hydroxysteroid Dehydrogenases / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D002822:Choriocarcinoma / Q000201:enzymology* / Q000473:pathology*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000201:enzymology* / Q000473:pathology*
D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D015067:17-Hydroxysteroid Dehydrogenases