title: Cross-talk between alpha1D-adrenoceptors and transient receptor potential vanilloid type 1 triggers prostate cancer cell proliferation.
abstract: BACKGROUND
  There is evidence that calcium (Ca(2+)) increases the proliferation of human advanced prostate cancer (PCa) cells but the ion channels involved are not fully understood. Here, we investigated the correlation between alpha(1D)-adrenergic receptor (alpha(1D)-AR) and the transient receptor potential vanilloid type 1 (TRPV1) expression levels in human PCa tissues and evaluated the ability of alpha(1D)-AR to cross-talk with TRPV1 in PCa cell lines.
  
  
  METHODS
  The expression of alpha1D-AR and TRPV1 was examined in human PCa tissues by quantitative RT-PCR and in PCa cell lines (DU145, PC3 and LNCaP) by cytofluorimetry. Moreover, alpha(1D)-AR and TRPV1 colocalization was investigated by confocal microscopy in PCa cell lines and by fluorescence microscopy in benign prostate hyperplasia (BPH) and PCa tissues. Cell proliferation was assessed by BrdU incorporation. Alpha(1D)-AR/TRPV1 knockdown was obtained using siRNA transfection. Signalling pathways were evaluated by measurement of extracellular acidification rate, Ca(2+) flux, IP3 production, western blot and MTT assay.
  
  
  RESULTS
  The levels of the alpha(1D)-AR and TRPV1 mRNAs are increased in PCa compared to BPH specimens and a high correlation between alpha(1D)-AR and TRPV1 expression levels was found. Moreover, alpha(1D)-AR and TRPV1 are co-expressed in prostate cancer cell lines and specimens. Noradrenaline (NA) induced an alpha(1D)-AR- and TRPV1-dependent protons release and Ca(2+) flux in PC3 cell lines; NA by triggering the activation of phospholipase C (PLC), protein kinase C (PKC) and extracellular signal-regulated kinase 1/2 (ERK1/2) pathways stimulated PC3 cell proliferation, that was completely inhibited by clopenphendioxan (WS433) and capsazepine (CPZ) combination or by alpha(1D)-AR/TRPV1 double knockdown.
  
  
  CONCLUSIONS
  We demonstrate a cross-talk between alpha1D-AR and TRPV1, that is involved in the control of PC3 cell proliferation. These data strongly support for a putative novel pharmacological approach in the treatment of PCa by targeting both alpha1D-AR and TRPV1 channels.
authors.forename: Maria Beatrice
authors.initials: MB
authors.lastname: Morelli
authors.affiliation: School of Biosciences and Veterinary Medicine, University of Camerino, Camerino 62032, Italy. consuelo.amantini@unicam.it.
authors.forename: Consuelo
authors.initials: C
authors.lastname: Amantini
authors.forename: Massimo
authors.initials: M
authors.lastname: Nabissi
authors.forename: Sonia
authors.initials: S
authors.lastname: Liberati
authors.forename: Claudio
authors.initials: C
authors.lastname: Cardinali
authors.forename: Valerio
authors.initials: V
authors.lastname: Farfariello
authors.forename: Daniele
authors.initials: D
authors.lastname: Tomassoni
authors.forename: Wilma
authors.initials: W
authors.lastname: Quaglia
authors.forename: Alessandro
authors.initials: A
authors.lastname: Piergentili
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bonifazi
authors.forename: Fabio
authors.initials: F
authors.lastname: Del Bello
authors.forename: Matteo
authors.initials: M
authors.lastname: Santoni
authors.forename: Gabriele
authors.initials: G
authors.lastname: Mammana
authors.forename: Lucilla
authors.initials: L
authors.lastname: Servi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Filosa
authors.forename: Angela
authors.initials: A
authors.lastname: Gismondi
authors.forename: Giorgio
authors.initials: G
authors.lastname: Santoni
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015870:Gene Expression; D006801:Humans; D008297:Male; D009638:Norepinephrine / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D011485:Protein Binding; D021381:Protein Transport; D018340:Receptors, Adrenergic, alpha-1 / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D050916:TRPV Cation Channels / Q000235:genetics* / Q000378:metabolism*
chemical_list: C547719:ADRA1D protein, human; D018340:Receptors, Adrenergic, alpha-1; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human; D009638:Norepinephrine