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Dual regulation of proliferation and growth arrest in prostatic stromal cells by transforming growth factor-beta1
Wei Zhou1, Irwin Park, Michael Pins
1Department of Urology, Northwestern University, Feinberg School of Medicine, Chicago, Illinois 60611, USA.
Abstract:
In a preliminary study, we observed that TGF-beta1 induced both proliferation and growth arrest in prostatic stromal cells, depending on the concentration of TGF-beta1 used in the culture medium. In this study, we explored possible mechanisms of this dual effect of TGF-beta. Primary cultures of prostatic stromal cells, established from clinical surgical specimens and treated with low doses of TGF-beta1 (0.001-0.01 ng/ml), resulted in an increase in cell proliferation. The addition of neutralizing antibody against platelet-derived growth factor (PDGF)-BB, but not anti-PDGF-AA, abrogated this stimulatory effect of TGF-beta1. TGF-beta1 treatment resulted in a dose-related increase in PDGF-BB production as measured by ELISA. Cells underwent growth arrest at high concentrations of TGF-beta1 (1.0 and 10 ng/ml). An inhibitor of cyclin-dependent kinase (cdk), p15INK4b, was up-regulated at both transcript and protein levels in these cultures by TGF-beta1 in a dose-related manner as determined by RT-PCR and Western blot analysis. The transcript, but not the protein, for another cdk inhibitor, p21Cip1, was up-regulated with treatment of TGF-beta1 to these cells. Levels of other cdk inhibitors, such as p16INK4a and p27Kip1, were constitutively expressed in prostatic stromal cells and were not significantly affected by TGF-beta1 treatment. Finally, the growth arrest effect of TGF-beta1 was abrogated when antisense oligonucleotides to p15INH4b, but not p21Cip1, were added to the culture medium. These data indicate that the dual effect of TGF-beta1 is mediated, at least, by up-regulation of PDGF-BB and p15INK4b, respectively.
Insights
Transforming growth factor-beta1 (TGF-beta1) uniquely influences prostatic stromal cells, promoting proliferation at low doses via platelet-derived growth factor-BB (PDGF-BB) and growth arrest at high doses through p15INK4b upregulation.
Area of Science:
- Cell Biology
- Molecular Biology
- Prostate Cancer Research
Background:
- Prostatic stromal cells play a crucial role in prostate tissue homeostasis.
- Transforming growth factor-beta1 (TGF-beta1) exhibits complex regulatory functions in various cell types.
- Previous observations indicated a dual effect of TGF-beta1 on prostatic stromal cell proliferation and growth arrest.
Purpose of the Study:
- To elucidate the underlying mechanisms of TGF-beta1's dual effect on prostatic stromal cells.
- To investigate the role of platelet-derived growth factor (PDGF) in TGF-beta1-induced proliferation.
- To identify the specific cyclin-dependent kinase (cdk) inhibitors involved in TGF-beta1-mediated growth arrest.
Main Methods:
- Primary cultures of human prostatic stromal cells were established.
- Cells were treated with varying concentrations of TGF-beta1.
- Enzyme-linked immunosorbent assay (ELISA) measured PDGF-BB production.
- Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyzed cdk inhibitor expression.
- Neutralizing antibodies and antisense oligonucleotides were used to probe signaling pathways.
Main Results:
- Low doses of TGF-beta1 (0.001-0.01 ng/ml) stimulated prostatic stromal cell proliferation, an effect abrogated by anti-PDGF-BB but not anti-PDGF-AA antibodies.
- TGF-beta1 treatment led to a dose-dependent increase in PDGF-BB production.
- High doses of TGF-beta1 (1.0 and 10 ng/ml) induced growth arrest.
- TGF-beta1 upregulated both transcript and protein levels of the cdk inhibitor p15INK4b in a dose-dependent manner.
- The growth arrest induced by TGF-beta1 was reversed by antisense oligonucleotides targeting p15INK4b, but not p21Cip1.
Conclusions:
- TGF-beta1 exerts a concentration-dependent dual effect on prostatic stromal cells.
- TGF-beta1-induced proliferation is mediated by the upregulation of PDGF-BB.
- TGF-beta1-induced growth arrest is primarily mediated by the upregulation of the cdk inhibitor p15INK4b.
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