胆管癌（CCA）是一种预后不佳的胆管恶性肿瘤。本研究系统地研究了核糖体蛋白 S6 (RPS6) 基因的作用，该基因依赖于 CCA。我们发现 CCA 组织中 RPS6 上调与不良预后相关。功能研究表明，RPS6 表达的改变、功能的获得和丧失都可能影响 CCA 细胞的增殖。在异种移植肿瘤模型中，RPS6 过表达会增强致瘤性，而 RPS6 沉默则会降低致瘤性。使用 RNA-seq 和蛋白质组学进行整合分析，通过影响细胞周期，尤其是 DNA 复制，阐明了 RPS6 耗竭的下游信号通路。免疫沉淀和质谱分析已鉴定出许多与 RPS6 相关的剪接体复合蛋白。转录组分析显示，RPS6 影响许多选择性剪接 (AS) 事件，并与 RNA 免疫沉淀测序相结合，揭示微型染色体维持复合物成分 7 (MCM7) 与 RPS6 结合，调节其 AS 并增加 CCA 的致癌活性。使用体内磷酸二酰胺吗啉寡聚物 (V-PMO) 靶向 RPS6 可显着抑制 CCA 细胞、患者来源的类器官和皮下异种移植肿瘤的生长。总而言之，数据表明 RPS6 是 CCA 的致癌调节因子，并且 RPS6-V-PMO 可以重新定位为治疗 CCA 的有前途的策略。© 2024 作者。

Cholangiocarcinoma (CCA) is a bile duct malignancy with a dismal prognosis. This study systematically investigated the role of the ribosomal protein S6 (RPS6) gene, which is dependent in CCA. We found that RPS6 upregulation in CCA tissues was correlated with a poor prognosis. Functional investigations have shown that alterations in RPS6 expression, both gain- and loss-of function could affect the proliferation of CCA cells. In xenograft tumor models, RPS6 overexpression enhances tumorigenicity, whereas RPS6 silencing reduces it. Integration analysis using RNA-seq and proteomics elucidated downstream signaling pathways of RPS6 depletion by affecting the cell cycle, especially DNA replication. Immunoprecipitation followed by mass spectrometry has identified numerous spliceosome complex proteins associated with RPS6. Transcriptomic profiling revealed that RPS6 affects numerous alternative splicing (AS) events, and combined with RNA immunoprecipitation sequencing, revealed that minichromosome maintenance complex component 7 (MCM7) binds to RPS6, which regulates its AS and increases oncogenic activity in CCA. Targeting RPS6 with vivo phosphorodiamidate morpholino oligomer (V-PMO) significantly inhibited the growth of CCA cells, patient-derived organoids, and subcutaneous xenograft tumor. Taken together, the data demonstrate that RPS6 is an oncogenic regulator in CCA and that RPS6-V-PMO could be repositioned as a promising strategy for treating CCA.© 2024 The Authors.