泛素蛋白酶体系统（UPS）的功能障碍促进了肝细胞癌（HCC）的恶性进展。虽然已经提出以 UPS 为目标进行 HCC 治疗，但确定有效的靶点一直具有挑战性。在这项研究中，我们对针对UPS相关WD40重复（WDR）蛋白的siRNA文库进行了重点筛选，发现沉默WDR20（一种去泛素化酶激活因子）可以选择性抑制HCC细胞的增殖，而不影响正常肝细胞。此外，在异种移植、睡美人转座子/转座酶、水动力尾静脉注射诱导的 HCC 模型以及 Alb-Cre /MYC HCC 转基因小鼠模型中，WDR20 表达的下调诱导 HCC 细胞衰老并抑制肿瘤进展。从机制上讲，我们发现 WDR20 沉默扰乱了 c-Myc 的蛋白质稳定性，同时协调了 USP12/46 介导的 c-Myc 去泛素化，从而促进了 CDKN1A 的转录激活。进一步研究显示，88 个 HCC 临床样本的组织微阵列中 WDR20 和 c-Myc 呈阳性共表达。通过使用来自 HCC 个体的三种患者来源的类器官，我们验证了 WDR20 沉默后 c-Myc 表达的减少以及衰老和生长抑制的显着诱导。这项研究不仅揭示了WDR20的生物学功能并阐明了其对HCC细胞衰老负调控的分子机制，而且还强调了WDR20作为HCC治疗有前景的靶点的潜力。

The dysfunction of the ubiquitin-proteasome system (UPS) facilitates the malignant progression of hepatocellular carcinoma (HCC). While targeting the UPS for HCC therapy has been proposed, identifying effective targets has been challenging. In this study, we conducted a focused screen of siRNA libraries targeting UPS-related WD40 repeat (WDR) proteins and found that silencing WDR20, a deubiquitinating enzyme activating factor, selectively inhibited the proliferation of HCC cells without affecting normal hepatocytes. Moreover, the downregulation of WDR20 expression induced HCC cellular senescence and suppressed tumor progression in xenograft, sleeping beauty transposon/transposase, and hydrodynamic tail vein injection-induced HCC models, and Alb-Cre+/MYC+ HCC transgenic mouse models. Mechanistically, we found that WDR20 silencing disturbed the protein stability of c-Myc, orchestrating the simultaneous USP12/46-mediated deubiquitination of c-Myc, thereby promoting the transcriptional activation of CDKN1A. Further investigation revealed a positive coexpression of WDR20 and c-Myc in a tissue microarray with 88 HCC clinical samples. By employing three patient-derived organoids from individuals with HCC, we have validated the decrease in c-Myc expression and the significant induction of senescence and growth inhibition following silencing of WDR20. This study not only uncovers the biological function of WDR20 and elucidates the molecular mechanism underlying its negative regulation of HCC cellular senescence but also highlight the potential of WDR20 as a promising target for HCC therapy.