17β-hydroxysteroid dehydrogenase types 1 and 2 : expression and activities in various tissues and cell lines and effect of the type 1 enzyme on estrogen-dependent growth of breast cancer cells
1University of Oulu, Biocenter Oulu
2University of Oulu, Faculty of Medicine, WHO Collaborating Centre for Research on Reproductive Health
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|Persistent link:|| http://urn.fi/urn:isbn:9514254163
|Publish Date:|| 1999-10-15
|Thesis type:||Doctoral Dissertation
|Defence Note:||Academic Dissertation to be presented with the assent of the Faculty of Medicine, University of Oulu, for public discussion in Auditorium 9 of the University Hospital of Oulu, on November 19th, 1999, at 12 noon.
Docent Helena Autio-Harmainen
Docent Jorma J. Palvimo
17β-Hydroxysteroid dehydrogenases (17HSDs) catalyze the reactions between 17-hydroxy and 17-keto steroids. In the present study, the enzyme activities and tissue distribution of 17HSD type 1, type 2 and type 4 were characterized. Furthermore, the role of 17HSD type 1 in estrogen-dependent growth was studied in MCF-7 breast cancer cells which were stably transfected with type 1 cDNA.
Endogenous oxidative 17HSD activity found in COS-m6 monkey kidney cells was first compared with that of human placental 17HSD. Cultured COS-m6 cells exclusively possessed oxidative 17HSD activity, converting estradiol (E2) to less active estrone (E1). When placental 17HSD was transfected into these cells, highly reductive activity appeared. The 17HSD enzyme in COS-m6 cells also catalyzed the conversion of testosterone to androstenedione, whereas the placental enzyme was estrogen-specific. These results further proved the existence of different 17HSD isoenzymes.
The enzymatic properties and cell- and tissue-specific expression of 17HSD type 1, type 2 and oxidative type 4 were further characterized. The data confirmed that in cultured cells the direction of 17HSD activity is determined by the expression of different isoenzymes and not by the intracellular environment. In addition, the 17HSD type 1 gene expresses two mRNA signals, 1.3 kb and 2.3 kb in size. The expression of 1.3 kb mRNA, but not 2.3 kb mRNA was related to enzyme concentration in all the cell types studied. The type 1 enzyme was expressed in the placenta, ovary and in some breast cancer specimens and in the cell lines originated from these tissues. 17HSD type 2 was more widely expressed in both steroidogenic and in target tissues of steroid action. 17HSD type 4 was expressed in almost all cell lines and in all tissues studied, but no correlation with 17HSD activity was detected. These results suggest that 17HSD type 1 is involved in E2 production in females and 17HSD type 2 is responsible for inactivation of sex steroids. However, the oxidation of 17β-hydroxysteroids seems not to be the primary activity of 17HSD type 4.
The mRNAs for 17HSD type 1, type 2 and type 4 were found to be expressed in human mammary epithelial cells. In breast tissue samples both 17HSD type 1 and type 2 were detected by in situ hybridization. Despite the presence of 17HSD type 1 mRNA in human mammary epithelial cells, only oxidative 17HSD activity was detected. The reason for the lack of reductive activity is not yet known.
Finally, MCF-7 breast cancer cells were stably transfected with 17HSD type 1 cDNA in order to study the effect of 17HSD type 1 on estrogen-dependent growth. In wild type MCF-7 cells, very low 17HSD activity was detected and E1 did not have any effect on cell growth. In the cells expressing 17HSD type 1, E1 was rapidly converted to E2. Hence in these cells E1 had a similar growth-promoting effect as E2 as a result of the action of 17HSD type 1. The presence of 17HSD type 1 in breast cancer cells may thus be an important factor regulating estrogen exposure and the estrogen-responsive growth of breast cancer tissue.
Acta Universitatis Ouluensis. D, Medica
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