Norepinephrine activates P44 and P42 MAPK in human prostate stromal and smooth muscle cells but not in epithelial cells.
2003
BACKGROUND
In vascular smooth muscle cells, α1-adrenergic stimulation increases DNA synthesis and cell proliferation via activation of p44/42 (ERK1/2) MAPK. We examined whether norepinephrine (NE) activates MAPK and stimulates the proliferation of prostatic epithelial and non-epithelial cells.
METHODS
Human prostatic epithelial cells, stromal cells, and smooth muscle cells were purchased from BioWhittaker (Walkersville, MD). After reaching a semi-confluent condition, the cells were cultured in RPMI-1640 without serum for 1 day. At 10 min after adding NE (10−6 or 10−7 M) to the medium, the cells were collected. Cell lysate was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) followed by Western blot using anti-phospho-p44/42 and anti-p44/42 antibodies. The activation of p44/42 was estimated by the ratio of phospho-p44/42 to total p44/42. Cell proliferation was evaluated by 3H-thymidine uptake assay. After reaching a semi-confluent condition, the cells were cultured in RPMI-1640 containing 0.5% FCS with or without NE (10−6 or 10−7 M) for 16 hr followed by a 3H-thymidine uptake period (24 hr).
RESULTS
P44/42 MAPK was significantly activated by NE in non-epithelial cells (stromal cells and smooth muscle cells) while not in epithelial cells. The uptake of 3H-thymidine was significantly increased by NE in both non-epithelial cells, which was inhibited by α1-adrenoceptor antagonists.
CONCLUSIONS
These results suggest that NE may stimulate the proliferation of non-epithelial prostatic cells, which may be involved in the pathogenesis of BPH. Prostate 56: 313–318, 2003. © 2003 Wiley-Liss, Inc.
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