title: SMYD3 as an oncogenic driver in prostate cancer by stimulation of androgen receptor transcription.
abstract: BACKGROUND
  Androgen receptor (AR) is critical for prostate tumorigenesis and is frequently overexpressed during prostate cancer (PC) progression. However, few studies have addressed the epigenetic regulation of AR expression.
  
  
  METHODS
  We analyzed SMYD3 expression in human PC with Western blot and immunohistochemistry. SMYD3 expression was knocked down using short hairpin RNA (shRNA) or small interfering RNA (siRNA). Cell proliferation, colony formation, and apoptosis analyses and xenograft transplantation were performed to evaluate the impact of SMYD3 depletion on PC cells. AR expression and promoter activity were determined using real-time quantitative polymerase chain reaction, western blot, and luciferase reporter assay. AR promoter association with Sp1, SMYD3, and histone modifications was assessed by chromatin immunoprecipitation. Differences in AR mRNA abundance and promoter activity were analyzed using Wilcoxon signed-rank tests, SMYD3 expression was analyzed using with Mann-Whitney U tests for unpaired samples, and tumor weight was analyzed with Student t test. All statistical tests were two-sided.
  
  
  RESULTS
  The upregulation of SMYD3 protein expression was observed in seven of eight prostate tumor specimens, compared with matched normal tissues. Immunohistochemical analysis showed a strong SMYD3 staining in the nuclei of PC tissues in eight of 25 (32%) cases and in the cytoplasm in 23 out of 25 (92%) cases, whereas benign prostate tissue exhibited weak immunostaining. Depletion of SMYD3 by siRNA or shRNA inhibited PC cell proliferation (72 hours relative to 24 hours: control shRNA vs SMYD3 shRNA 1: mean fold change = 2.76 vs 1.68; difference = 1.08; 95% confidence interval = 0.78 to 1.38, P < .001), colony formation, cell migration, invasion, and xenograft tumor formation. Two functional SMYD3-binding motifs were identified in the AR promoter region.
  
  
  CONCLUSIONS
  SMYD3 promotes prostate tumorigenesis and mediates epigenetic upregulation of AR expression.
authors.affiliation: Affiliations of authors: Department of Urology (CL, CW, KW, KY, JC, JL, HR, HL, ZX, YF), Department of General Surgery (LL, SH), and School of Nursing (LL), Shandong University Qilu Hospital, Jinan, Shandong, China; Central Research Laboratory of Shandong University Second Hospital, Jinan, Shandong, China (DX); Department of Urology, Peking University First Hospital, Beijing, China (QS); Department of Urology, Qingdao Municipal Hospital, Qingdao, China (XS); Department of Medicine, Division of Haematology and Centre for Molecular Medicine, Karolinska University Hospital Solna and Karolinska Institutet, Stockholm, Sweden (CW, DX).
authors.forename: Cheng
authors.initials: C
authors.lastname: Liu
authors.forename: Chang
authors.initials: C
authors.lastname: Wang
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Qi
authors.initials: Q
authors.lastname: Shen
authors.forename: Keqiang
authors.initials: K
authors.lastname: Yan
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Sun
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
authors.forename: Jikai
authors.initials: J
authors.lastname: Liu
authors.forename: Hongbo
authors.initials: H
authors.lastname: Ren
authors.forename: Hainan
authors.initials: H
authors.lastname: Liu
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Xu
authors.forename: Sanyuan
authors.initials: S
authors.lastname: Hu
authors.forename: Dawei
authors.initials: D
authors.lastname: Xu
authors.forename: Yidong
authors.initials: Y
authors.lastname: Fan
mesh_terms: D063646:Carcinogenesis; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D047369:Chromatin Immunoprecipitation; D044127:Epigenesis, Genetic; D015972:Gene Expression Regulation, Neoplastic; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D009361:Neoplasm Invasiveness; D011401:Promoter Regions, Genetic; D064129:Prostatic Neoplasms, Castration-Resistant / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D034741:RNA, Small Interfering / Q000494:pharmacology; D060888:Real-Time Polymerase Chain Reaction; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism; D012844:Sincalide / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D015854:Up-Regulation; D023041:Xenograft Model Antitumor Assays
chemical_list: C410798:AR protein, human; D034741:RNA, Small Interfering; D011944:Receptors, Androgen; D011495:Histone-Lysine N-Methyltransferase; C506278:SMYD3 protein, human; D012844:Sincalide