胆管癌（CCA）具有高度异质性和极度恶性的特点，预后较差。双皮质素样激酶 1 (DCLK1) 促进多种恶性肿瘤的进展。以 DCLK1 或其相关调节途径为靶点可以预防多种恶性肿瘤的产生和恶化。然而，DCLK1 在 CCA 进展中的作用及其分子机制仍不清楚。因此，我们旨在研究DCLK1是否以及如何促进CCA进展。使用免疫组织化学（IHC）检测CCA患者中DCLK1的表达。我们建立了 DCLK1 敲除和 DCLK1 过表达细胞系，用于集落形成实验和 Transwell 实验，以探索 DCLK1 的促肿瘤作用。 RT-PCR、Western blot 和多重荧光染色用于评估 DCLK1 和上皮间质转化 (EMT) 标记物之间的关联。通过 RNA 测序和生物信息学分析来确定 DCLK1 调节 CCA 进展和 EMT 程序的潜在机制。DCLK1 在 CCA 组织中过度表达，并与不良预后相关。 DCLK1过表达促进CCA细胞侵袭、迁移和增殖，而DCLK1敲低则逆转CCA细胞的恶性倾向，这一点在体内和体外均得到证实。此外，我们证明 DCLK1 与 EMT 程序的进展密切相关，其中包括间充质标记物的过度表达和上皮标记物的下调。对于其潜在机制，我们提出PI3K/AKT/mTOR通路是DCLK1在肿瘤进展和EMT程序发生中发挥作用的关键过程。当给予PI3K/AKT/mTOR通路抑制剂LY294002时，肿瘤的增殖、迁移和侵袭能力被极大抑制，EMT过程普遍逆转。DCLK1通过PI3K促进CCA细胞的恶性生物学行为/AKT/mTOR 途径。对于高水平表达 DCLK1 的胆管癌个体，PI3K/AKT/mTOR 信号通路抑制剂可能是一种有效的治疗方法。© 2024。作者。

Cholangiocarcinoma (CCA), characterized by high heterogeneity and extreme malignancy, has a poor prognosis. Doublecortin-like kinase 1 (DCLK1) promotes a variety of malignant cancers in their progression. Targeting DCLK1 or its associated regulatory pathways can prevent the generation and deterioration of several malignancies. However, the role of DCLK1 in CCA progression and its molecular mechanisms remain unknown. Therefore, we aimed to investigate whether and how DCLK1 contributes to CCA progression.The expression of DCLK1 in CCA patients was detected using Immunohistochemistry (IHC). We established DCLK1 knockout and DCLK1 overexpression cell lines for Colony Formation Assay and Transwell experiments to explore the tumor-promoting role of DCLK1. RT-PCR, Western blot and multiple fluorescent staining were used to assess the association between DCLK1 and epithelial-mesenchymal transition (EMT) markers. RNA sequencing and bioinformatics analysis were performed to identify the underlying mechanisms by which DCLK1 regulates CCA progression and the EMT program.DCLK1 was overexpressed in CCA tissues and was associated with poor prognosis. DCLK1 overexpression facilitated CCA cell invasion, migration, and proliferation, whereas DCLK1 knockdown reversed the malignant tendencies of CCA cells, which had been confirmed both in vivo and in vitro. Furthermore, we demonstrated that DCLK1 was substantially linked to the advancement of the EMT program, which included the overexpression of mesenchymal markers and the downregulation of epithelial markers. For the underlying mechanism, we proposed that the PI3K/AKT/mTOR pathway is the key process for the role of DCLK1 in tumor progression and the occurrence of the EMT program. When administered with LY294002, an inhibitor of the PI3K/AKT/mTOR pathway, the tumor's ability to proliferate, migrate, and invade was greatly suppressed, and the EMT process was generally reversed.DCLK1 facilitates the malignant biological behavior of CCA cells through the PI3K/AKT/mTOR pathway. In individuals with cholangiocarcinoma who express DCLK1 at high levels, inhibitors of the PI3K/AKT/mTOR signaling pathway may be an effective therapeutic approach.© 2024. The Author(s).