目标:CLSPN,S期检查点响应DNA复制应激的关键组成部分,与多种肿瘤类型的发病机理有关。肝细胞癌(HCC)发病率的上升对全球公共卫生构成了重大挑战。尽管如此,CLSPN在HCC发展中的具体功能仍然知之甚少。
方法:我们系统地评估了CLSPN的表达,肝癌患者的预后和免疫浸润,并通过使用公共数据库确定竞争内源性RNA(ceRNA)网络。RT-qPCR,westernblot,CCK8Transwell,流式细胞术,动物实验,蛋白酶体抑制实验,采用Co-IP法和质谱联用技术探讨其生物学功能,CLSPN在HCC中的转录后修饰和潜在分子机制。
结果:我们验证了CLSPN的表达,其高表达是肝癌的独立预后因素。CLSPN的表达也与HCC的免疫微环境有关。CLSPN沉默抑制增殖,迁移,肝癌细胞的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受泛素化的影响,参与Wnt/β-catenin通路调节HCC进展。
结论:这是第一次全面发现和鉴定表达,预后,免疫疗法,RNA调节因子,转录后修饰,和CLSPN在HCC中的分子机制。这些新颖的见解有可能加快肝癌患者个性化治疗策略和转化医学方法的发展。
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.