Prostasin regulates iNOS and cyclin D1 expression by modulating protease-activated receptor-2 signaling in prostate epithelial cells.

Abstract

BACKGROUND:

Prostasin is down-regulated during inflammation and in invasive cancers, and plays a role in regulation of inflammatory gene expression and invasion.

METHODS:

We used the human benign prostatic hyperplasia cell line BPH-1 to investigate gene expression changes associated with siRNA-mediated loss of prostasin expression. Quantitative PCR and/or western blotting were used to evaluate the expression changes of iNOS, ICAM-1, cyclin D1, IL-6, and IL-8. Gene expression changes were also evaluated in the presence of a PAR-2 antagonist. The PC-3 human prostate cancer cell line was used for evaluation of gene expression in response to prostasin re-expression.

RESULTS:

Prostasin silencing in BPH-1 was associated with up-regulation of iNOS, ICAM-1, IL-6, and IL-8, and down-regulation of cyclin D1; as well as reduced proliferation and invasion. The iNOS up-regulation and cyclin D1 down-regulation associated with prostasin silencing were inhibited by a PAR-2 antagonist. Re-expression of prostasin, a serine active-site mutant, and a GPI-anchor-free mutant, in the PC-3 cells resulted in PAR-2 and cyclin D1 transcription up-regulation. Transcription up-regulation of IL-6 and IL-8 was associated with re-expression of the serine active-site mutant prostasin in the PC-3 cells. Transcription up-regulation of IL-8, but to a lesser extent, was also observed in PC-3 cells expressing the wild-type prostasin. Expression of a serine protease active prostasin, GPI-anchored or anchor-free, prevented the IL-6 induction in response to PAR-2. The GPI-anchor-free prostasin also prevented the IL-8 induction.

CONCLUSIONS:

Prostasin plays a negative regulatory role on PAR-2-mediated signaling in prostate epithelial cells.

• Authors: Li-Mei Chen, Meghan L Hatfield, Ya-Yuan Fu, Karl X Chai
• Journal: The Prostate (Prostate) Vol. 69 Issue 16 Pg. 1790-801 (Dec 01 2009) ISSN: 1097-0045 [Electronic] United States
• PMID: 19670249 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2009 Wiley-Liss, Inc.

Chemical References

• Interleukin-6
• Interleukin-8
• RNA, Small Interfering
• Receptor, PAR-2
• Intercellular Adhesion Molecule-1
• Cyclin D1
• NOS2 protein, human
• Nitric Oxide Synthase Type II
• Serine Endopeptidases
• prostasin

Topics

• Blotting, Western
• Cell Line
• Cell Proliferation (drug effects)
• Cyclin D1 (metabolism)
• Down-Regulation (drug effects)
• Epithelial Cells (metabolism)
• Humans
• Intercellular Adhesion Molecule-1 (metabolism)
• Interleukin-6 (metabolism)
• Interleukin-8 (metabolism)
• Male
• Nitric Oxide Synthase Type II (metabolism)
• Polymerase Chain Reaction
• Prostate (metabolism, pathology)
• Prostatic Hyperplasia (metabolism, pathology)
• RNA, Small Interfering (pharmacology)
• Receptor, PAR-2 (metabolism)
• Serine Endopeptidases (genetics, metabolism)
• Signal Transduction
• Up-Regulation (drug effects)