SMYD3 as an Oncogenic Driver in Prostate Cancer by Stimulation of Androgen Receptor Transcription
SMYD3 as an Oncogenic Driver in Prostate Cancer by Stimulation of Androgen Receptor Transcription
复制标题
SMYD3 通过刺激雄激素受体转录作为前列腺癌的致癌驱动因素
DOI：
10.1093/jnci/djt304
复制
发表时间：
2013-11-01
影响因子：
10.3
通讯作者：
Fan, Yidong
中科院分区：
文献类型：
--
作者：
Liu, Cheng;Wang, Chang;Fan, Yidong
雄激素受体（AR）对前列腺肿瘤发生至关重要，并且在前列腺癌（PC）进展期间经常过表达。然而，很少有研究涉及AR表达的表观遗传调控。使用短发夹RNA（shRNA）或小干扰RNA（siRNA）敲低SMYD 3表达。进行细胞增殖、集落形成和凋亡分析以及异种移植物移植以评估SMYD 3耗竭对PC细胞的影响。采用实时定量聚合酶链反应、蛋白质印迹和荧光素酶报告基因分析测定AR表达和启动子活性。AR启动子与Sp1，SMYD 3和组蛋白修饰的关联通过染色质免疫沉淀进行评估。使用Wilcoxon符号秩检验分析AR mRNA丰度和启动子活性的差异，使用MannWhitney U检验分析未配对样品的SMYD 3表达，并使用Student t检验分析肿瘤重量。所有的统计学检验都是双侧的，与匹配的正常组织相比，在8个前列腺肿瘤标本中有7个观察到SMYD 3蛋白表达上调。免疫组化分析显示，25例中有8例（32%）PC组织的细胞核和25例中的23例（92%）细胞质中有强SMYD 3染色，而良性前列腺组织的免疫染色较弱。通过siRNA或shRNA消耗SMYD 3抑制PC细胞增殖（72小时相对于24小时：对照shRNA vs SMYD 3 shRNA 1：平均倍数变化2.76 vs 1.68;差异1.08; 95%置信区间0.78至1.38，P < .001）、集落形成、细胞迁移、侵袭和异种移植肿瘤形成。在AR启动子区域发现了两个SMYD 3结合基序，SMYD 3促进前列腺肿瘤发生并介导AR表达的表观遗传上调。
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.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 MannWhitney U tests for unpaired samples, and tumor weight was analyzed with Student t test. All statistical tests were two-sided.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.SMYD3 promotes prostate tumorigenesis and mediates epigenetic upregulation of AR expression.