肝内胆管癌 (iCCA) 是一种高度侵袭性的外周胆管癌症，可通过肿瘤内大量的癌症干细胞 (CSC) 来识别。虽然 iCCA 中的 CSC 标记物定义明确，但该亚群的分子脆弱性仍然难以捉摸。96 孔三维 (3D) 肿瘤球培养物改编自完善的 CSC 模型，通过基因表达分析验证了 CSC 标记物。然后进行激酶库筛选，以揭示潜在的致癌脆弱途径。利用RNA干扰稳定沉默三种iCCA细胞系中的候选基因，并评估其对iCCA细胞增殖和肿瘤球形成效率（TFE）的影响。激酶抑制剂库筛选确定了对肿瘤球活力至关重要的前50种激酶抑制剂，其中有11种抑制剂靶向 IGF-1R/PI3K/AKT 轴。对最热门抑制剂的进一步剂量依赖性分析证实 IGF-1R 是候选分子。 IGF-1R 稳定沉默后，所有三种 iCCA 细胞系均表现出 AKT 激活减少、增殖受阻和 TFE 减少，表明 CSC 亚群减少。IGF-1R 在维持 iCCA 干细胞群中发挥着关键作用。我们的数据强调IGF-1R 作为 iCCA 预后标志物和消除其 CSC 亚群的治疗靶点的潜在用途。版权所有 © 2024 Elsevier B.V. 保留所有权利。

Intrahepatic cholangiocarcinoma (iCCA) is a highly aggressive cancer of the peripheral bile ducts and is recognized by the abundance of cancer stem-like cells (CSCs) within the tumor mass. While CSC markers in iCCA are well-defined, the molecular vulnerabilities of this subpopulation remain elusive.The 96-well, three dimensional (3D) tumorsphere culture was adapted from a well-established CSC model, validated for CSC markers through gene expression analysis. Kinase library screening was then conducted to reveal potential oncogenic vulnerable pathways. RNA interference was utilized to stably silence the candidate gene in three iCCA cell lines and its impact on iCCA cell proliferation and tumorsphere formation efficiency (TFE) was evaluated.Kinase inhibitor library screening identified the top 50 kinase inhibitors crucial for tumorsphere viability, with 11 inhibitors targeting the IGF-1R/PI3K/AKT axis. Further dose-dependent analysis of the top 'hit' inhibitors confirmed IGF-1R as the candidate molecule. Upon stably silencing of IGF-1R, all three iCCA cell lines exhibited decreased AKT activation, impeded proliferation and reduced TFE, indicating a decline in CSC subpopulations.IGF-1R plays a critical role in maintaining iCCA-stem like cell populations.Our data highlight the potential utility of IGF-1R as a prognostic marker of iCCA and a therapeutic target for eliminating its CSC subpopulation.Copyright © 2024 Elsevier B.V. All rights reserved.