Cheng Liu1, Chang Wang, Kun Wang, Li Liu, Qi Shen, Keqiang Yan, Xiaoqing Sun, Jie Chen, Jikai Liu, Hongbo Ren, Hainan Liu, Zhonghua Xu, Sanyuan Hu, Dawei Xu, Yidong Fan. 1. 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).
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.
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: Christopher Tracy; Junco S Warren; Marta Szulik; Li Wang; June Garcia; Aman Makaju; Kristi Russell; Mickey Miller; Sarah Franklin Journal: Curr Opin Physiol Date: 2017-12-13
Authors: Lorna H Mitchell; P Ann Boriack-Sjodin; Sherri Smith; Michael Thomenius; Nathalie Rioux; Michael Munchhof; James E Mills; Christine Klaus; Jennifer Totman; Thomas V Riera; Alejandra Raimondi; Suzanne L Jacques; Kip West; Megan Foley; Nigel J Waters; Kevin W Kuntz; Tim J Wigle; Margaret Porter Scott; Robert A Copeland; Jesse J Smith; Richard Chesworth Journal: ACS Med Chem Lett Date: 2015-08-27 Impact factor: 4.345
Authors: Emeli M Nilsson; Kristian B Laursen; Jonathan Whitchurch; Andrew McWilliam; Niels Ødum; Jenny L Persson; David M Heery; Lorraine J Gudas; Nigel P Mongan Journal: Oncotarget Date: 2015-11-03