title: Effects of the dual 5 alpha-reductase inhibitor dutasteride on apoptosis in primary cultures of prostate cancer epithelial cells and cell lines.
abstract: BACKGROUND
  The profound reduction in serum dihydrotestosterone (DHT) observed with the dual 5 alpha-reductase inhibitor (5ARI) dutasteride makes it an attractive agent for prostate cancer therapy. The objective of the current study was to determine whether dutasteride would induce apoptosis in a range of prostate epithelial cell lines and primary cultures.
  
  
  METHODS
  Both human prostate androgen-sensitive cell lines (PwR-1E, PNT-2, LNCaP, and PC3[AR2]) and an androgen-independent cell line (PC-3) were grown to confluence. Primary epithelial cells extracted from fresh prostate cancer radical prostatectomy specimens also were grown to confluence under optimal conditions. Total cellular protein was extracted to confirm cytokeratin 18 and antihuman alpha-methylacyl-CoA racemase (AMACR) expression of the primary cells. Apoptosis was assessed by propidium iodide DNA staining and flow cytometry after 24 hours of culture in from 0 microM to 10 microM of dutasteride.
  
  
  RESULTS
  Dutasteride induced a dose-dependent increase in apoptosis in the androgen-sensitive prostate cell lines PwR-1E, PNT-2, and LNCaP and in the androgen receptor-expressing PC3(AR2) cell line. However, there was no significant apoptosis noted in the parental PC-3 cells. Of 16 primary epithelial cultures that were treated, 7 cultures were induced to undergo apoptosis, and 9 cultures were unresponsive. All primary cultures were positive for cytokeratin 18 expression, confirming their epithelial phenotype. Responder epithelial cells were positive for AMACR expression.
  
  
  CONCLUSIONS
  The results of the current study confirmed that dutasteride differentially induced apoptosis in a subset of prostate cell lines and primary prostate epithelial cells. Understanding the cellular phenotype may indicate susceptible cells.
authors.affiliation: University College Dublin School of Medicine and Medical Sciences, Mater Misericordiae University Hospital, Dublin, Ireland.
authors.forename: Ann
authors.initials: A
authors.lastname: Maria McCrohan
authors.forename: Colm
authors.initials: C
authors.lastname: Morrissey
authors.forename: Conor
authors.initials: C
authors.lastname: O'Keane
authors.forename: Niall
authors.initials: N
authors.lastname: Mulligan
authors.forename: Chanel
authors.initials: C
authors.lastname: Watson
authors.forename: James
authors.initials: J
authors.lastname: Smith
authors.forename: John M
authors.initials: JM
authors.lastname: Fitzpatrick
authors.forename: R William G
authors.initials: RW
authors.lastname: Watson
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001378:Azasteroids / Q000494:pharmacology*; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D042944:Cholestenone 5 alpha-Reductase / Q000037:antagonists & inhibitors* / Q000502:physiology; D004273:DNA, Neoplasm / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D000068538:Dutasteride; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004847:Epithelial Cells / Q000737:chemistry / Q000187:drug effects* / Q000473:pathology / Q000502:physiology; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007633:Keratins / Q000032:analysis / Q000235:genetics; D008297:Male; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000473:pathology* / Q000503:physiopathology; D012333:RNA, Messenger / Q000032:analysis; D019998:Racemases and Epimerases / Q000032:analysis / Q000235:genetics; D011944:Receptors, Androgen / Q000032:analysis / Q000235:genetics
chemical_list: D001378:Azasteroids; D004273:DNA, Neoplasm; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D011944:Receptors, Androgen; D007633:Keratins; D042944:Cholestenone 5 alpha-Reductase; D019998:Racemases and Epimerases; C108352:alpha-methylacyl-CoA racemase; D000068538:Dutasteride