CLSPN, a critical component of the S-phase checkpoint in response to DNA replication stress, has been implicated in the pathogenesis of multiple tumor types. The rising incidence of hepatocellular carcinoma (HCC) poses a significant challenge to global public health. Despite this, the specific functions of CLSPN in the development of HCC remain poorly understood. We systematically evaluated the expression of CLSPN, prognosis and immune infiltration in patients with HCC and identified a competing endogenous RNA (ceRNA) network by using public database. The RT-qPCR, western blot, CCK8, transwell, flow cytometry, animal experiments, proteasome inhibition experiment, Co-IP assay and mass spectrometry were applied to explore its biological functions, post-transcriptional modifications and potential molecular mechanisms of CLSPN in HCC. We verified the expression of CLSPN, and its high expression is an independent prognostic factor in HCC. The expression of CLSPN is also associated with the immune microenvironment of HCC. CLSPN silencing inhibited the proliferation, migration, invasion and cell cycle progression of HCC cells. We established a PSMA3-AS1/hsa-miR-101-3p/CLSPN regulator axis in HCC. CLSPN was influenced by ubiquitination and was involved in the Wnt/β-catenin pathway to regulate HCC progression. It was the first time to comprehensively discover and identify the expression, prognosis, immunotherapy, RNAs regulator, posttranscriptional modification, and molecular mechanisms of CLSPN in HCC. These novel insights have the potential to expedite the development of personalized treatment strategies and translational medicine approaches for HCC patients.

中文翻译：

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CLSPN 如何揭开其对肝细胞癌的预后价值和潜在分子机制的神秘面纱：一项串扰研究

CLSPN 是响应 DNA 复制应激的 S 期检查点的关键组成部分，与多种肿瘤类型的发病机制有关。肝细胞癌（HCC）发病率的上升给全球公共卫生带来了重大挑战。尽管如此，CLSPN 在 HCC 发展中的具体功能仍然知之甚少。我们系统地评估了 HCC 患者 CLSPN 的表达、预后和免疫浸润，并利用公共数据库识别了竞争性内源性 RNA (ceRNA) 网络。应用RT-qPCR、western blot、CCK8、transwell、流式细胞术、动物实验、蛋白酶体抑制实验、Co-IP实验和质谱分析等手段探讨CLSPN在HCC中的生物学功能、转录后修饰及潜在分子机制。我们验证了CLSPN的表达，其高表达是HCC的独立预后因素。 CLSPN的表达还与HCC的免疫微环境有关。 CLSPN沉默抑制HCC细胞的增殖、迁移、侵袭和细胞周期进程。我们在 HCC 中建立了 PSMA3-AS1/hsa-miR-101-3p/CLSPN 调节轴。 CLSPN 受泛素化影响，参与 Wnt/β-catenin 通路调节 HCC 进展。首次全面发现和鉴定CLSPN在HCC中的表达、预后、免疫治疗、RNA调节、转录后修饰及分子机制。这些新颖的见解有可能加快针对 HCC 患者的个性化治疗策略和转化医学方法的开发。