Effect of beta-sitosterol on transforming growth factor-beta-1 expression and translocation protein kinase C alpha in human prostate stromal cells in vitro.
BACKGROUND
Today, plant extracts are widely used in the treatment of benign prostatic hyperplasia (BPH). However, the complete mode of action of the active substance, beta-sitosterol, is under investigation. The purpose of this study was to investigate the effect of beta-sitosterol on the expression of transforming growth factor beta 1 (TGF-beta1) and the activity of protein kinase C alpha (PKC-alpha) in primary prostate stromal cell cultures in vitro.


METHODS
Tissue samples for primary cell cultures were obtained from patients undergoing transurethral resection of the prostate (TURP). TGF-beta1 levels in stromal cell conditioned media following a culture with beta-sitosterol were detected in a TGF-beta1 specific ELISA assay. Following different incubation periods with beta-sitosterol, cells were lysed and fractionated into a Triton-soluble membrane fraction and a cytosol fraction. PKC-alpha protein was detected using immunoblot analysis.


RESULTS
Beta-sitosterol was able to induce the expression and secretion of TGF-beta1 significantly between 1.26- and 1.86-fold compared to a cholesterol and the nonsupplemented control in 6 of 8 individual cultures. The total amount of secreted TGF-beta1 varied in cells from different patients. Based on its presence in both membrane fraction and cytosol, PKC-alpha appeared to be constitutively expressed in stromal cells. In the absence of beta-sitosterol PKC-alpha was predominantly found in its membrane-associated active form. Following a culture with beta-sitosterol, a translocation of PKC-alpha from the membrane to the cytosol was observed. This effect was specific for beta-sitosterol as compared to cholesterol.


CONCLUSIONS
This study describes the effect of beta-sitosterol on the expression of a multifunctional growth factor (TGF-beta1) and the activity of PKC-alpha membrane in stromal cells of the human prostate in vitro.
Kassen
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Wissenschaftliches Labor der Urologischen Klinik der Ruhr-Universität Bochum, Deutschland. Annette.Kassen@ruhr-uni-bochum.de
Berges
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Senge
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D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008297:Male; D011467:Prostate / Q000166:cytology* / Q000201:enzymology*; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D012855:Sitosterols / Q000494:pharmacology*; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000096:biosynthesis*
D007527:Isoenzymes; D012855:Sitosterols; D016212:Transforming Growth Factor beta; C025473:gamma-sitosterol; C496628:PRKCA protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha