title: Zyflamend inhibits the expression and function of androgen receptor and acts synergistically with bicalutimide to inhibit prostate cancer cell growth.
abstract: BACKGROUND
  Interference of androgen receptor (AR) signaling is a target for prostate cancer (CaP) chemoprevention and treatment. We hypothesize that Zyflamend (ZYF) assert its anti-cancer effect by disrupting AR signaling. We also hypothesize that it may act synergistically with the anti-androgen bicalutimde to inhibit CaP cell growth.
  
  
  METHODS
  Western blotting, ELISA and reporter assays were done to test ZYF on AR signaling. Semi-quantitative RT-PCR and AR half-life were also examined. Potential synergism between ZYF and bicalutimide were tested via cytotoxicity, colony formation assays, flow cytometry, and Western blotting in the human CAP line, LNCaP and 22RV1.
  
  
  RESULTS
  ZYF reduced AR protein, mRNA and protein stability levels in LNCaPs. ZYF also reduced both full-length AR protein and truncated AR protein in the 22Rv1 cell line. Nkx3.1 and PSA were also reduced at the mRNA level. PSA promoter activity and secretion were lower after treatment of cells with ZYF. DHT induction of cell proliferation and AR responsiveness revealed reduction of AR, Nkx3.1, and PSA protein were demonstrated with ZYF treatment. Co-treatment with bicalutimide reducing cell growth, induced apoptosis, and reduced Bcl-2 and BclxL, caspase-3 and PARP. Co-treatment also reduced Nkx3.1 and PSA protein.
  
  
  CONCLUSIONS
  These data indicate that ZYF suppresses cell growth mediated by AR signaling, and suggests that the co-treatment with the anti-androgen bicalutimide and ZYF may be a promising approach for cancer therapy and may demonstrate the mechanism of action of ZYF.
authors.affiliation: Model Animal Research Center, MOE Key Laboratory of Model Animal for Disease Study, Nanjing University, Nanjing, China. yanjun@nicemice.cn
authors.forename: Jun
authors.initials: J
authors.lastname: Yan
authors.forename: Bingxian
authors.initials: B
authors.lastname: Xie
authors.forename: Jillian L
authors.initials: JL
authors.lastname: Capodice
authors.forename: Aaron E
authors.initials: AE
authors.lastname: Katz
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D059002:Androgen Receptor Antagonists / Q000494:pharmacology*; D000813:Anilides / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004357:Drug Synergism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D009570:Nitriles / Q000494:pharmacology*; D010936:Plant Extracts / Q000494:pharmacology*; D017430:Prostate-Specific Antigen / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D011944:Receptors, Androgen / Q000187:drug effects* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014105:Tosyl Compounds / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D059002:Androgen Receptor Antagonists; D000813:Anilides; D000970:Antineoplastic Agents; D018398:Homeodomain Proteins; C491178:NKX3-1 protein, human; D009570:Nitriles; D010936:Plant Extracts; D012333:RNA, Messenger; D011944:Receptors, Androgen; D014105:Tosyl Compounds; D014157:Transcription Factors; C519312:Zyflamend; C053541:bicalutamide; D017430:Prostate-Specific Antigen; D053148:Caspase 3