Department of Reproductive Medicine, Hexian Memorial Affiliated Hospital of Southern Medical University, Guangzhou, China.
Jiangxi Institute of Urology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China.
J Gene Med. 2023 Jul;25(7):e3498. doi: 10.1002/jgm.3498. Epub 2023 Mar 27.
WD repeat domain 3 (WDR3) is involved in tumor growth and proliferation, but its role in the pathological mechanism of prostate cancer (PCa) is still unclear.
WDR3 gene expression levels were obtained by analyzing databases and our clinical specimens. The expression levels of genes and proteins were determined by a real-time polymerase chain reaction, western blotting and immunohistochemistry, respectively. Cell-counting kit-8 assays were used to measure the proliferation of PCa cells. Cell transfection was used to investigate the role of WDR3 and USF2 in PCa. Fluorescence reporter and chromatin immunoprecipitation assays were used to detect USF2 binding to the promoter region of RASSF1A. Mouse experiments were used to confirm the mechanism in vivo.
By analyzing the database and our clinical specimens, we found that WDR3 expression was significantly increased in PCa tissues. Overexpression of WDR3 enhanced PCa cell proliferation, decreased cell apoptosis rate, increased spherical cell number and increased indicators of stem cell-like properties. However, these effects were reversed by WDR3 knockdown. WDR3 was negatively correlated with USF2, which was degraded by promoting ubiquitination of USF2, and USF2 interacted with promoter region-binding elements of RASSF1A to depress PCa stemness and growth. In vivo studies showed that WDR3 knockdown reduced tumor size and weight, reduced cell proliferation and enhanced cell apoptosis.
WDR3 ubiquitinated USF2 and inhibited its stability, whereas USF2 interacted with promoter region-binding elements of RASSF1A. USF2 transcriptionally activated RASSF1A, which inhibited the carcinogenic effect of WDR3 overexpression.
WD 重复结构域 3（WDR3）参与肿瘤生长和增殖，但它在前列腺癌（PCa）病理机制中的作用尚不清楚。
通过分析数据库和我们的临床标本获得 WDR3 基因表达水平。通过实时聚合酶链反应、western blot 和免疫组织化学分别测定基因和蛋白质的表达水平。细胞计数试剂盒-8 测定用于测量 PCa 细胞的增殖。细胞转染用于研究 WDR3 和 USF2 在 PCa 中的作用。荧光报告和染色质免疫沉淀测定用于检测 USF2 与 RASSF1A 启动子区域的结合。小鼠实验用于体内验证机制。
通过分析数据库和我们的临床标本，我们发现 WDR3 在 PCa 组织中的表达显著增加。WDR3 的过表达增强了 PCa 细胞的增殖，降低了细胞凋亡率，增加了球体细胞数量，并增加了干细胞样特性的指标。然而，WDR3 的敲低逆转了这些效应。WDR3 与 USF2 呈负相关，WDR3 通过促进 USF2 的泛素化降解 USF2，而 USF2 与 RASSF1A 启动子区域结合元件相互作用，抑制 PCa 干细胞特性和生长。体内研究表明，WDR3 的敲低降低了肿瘤的大小和重量，降低了细胞增殖并增强了细胞凋亡。
WDR3 泛素化 USF2 并抑制其稳定性，而 USF2 与 RASSF1A 启动子区域结合元件相互作用。USF2 转录激活 RASSF1A，抑制 WDR3 过表达的致癌作用。
