靶向PI3K/AKT/mTOR通路，為雙皮質(zhì)素樣激酶1高表達的膽管癌患者提供了一種治療新策略

2024年7月，首都醫(yī)科大學附屬朝陽醫(yī)院腫瘤科，清華大學醫(yī)學院基礎醫(yī)學部清華-北大生命科學中心免疫學研究所，首都醫(yī)科大學附屬朝陽醫(yī)院消化科（1Department of

Oncology, Beijing Chao Yang Hospital, Capital Medical University, 8 Gongren Tiyuchang Nanlu Road, Chaoyang Dist.2 Department of
Basic Medical Sciences, School of
Medicine, Tsinghua-Peking Center for
Life Sciences, Institute for
Immunology, Tsinghua University.3 Department of
Gastroenterology, Beijing Chao Yang Hospital, Capital Medical University, 8 Gongren Tiyuchang Nanlu Road, Chaoyang Dist.）Donglei Zhang研究團隊在《Journal of Cancer Research and Clinical Oncology》上發(fā)表論文：

“Targeting the PI3K/AKT/mTOR pathway offer a promising therapeutic strategy for cholangiocarcinoma patients with high doublecortin-like kinase 1 expression”

“靶向PI3K/AKT/mTOR通路，為雙皮質(zhì)素樣激酶1高表達的膽管癌患者提供了一種治療新策略”

Abstract：

Background 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.

Methods 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 fuorescent 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.

Results DCLK1 was overexpressed in CCA tissues and was associated with poor prognosis. DCLK1 overexpression facili

tated CCA cell invasion, migration, and proliferation, whereas DCLK1 knockdown reversed the malignant tendencies of

CCA cells, which had been confrmed both in vivo and in vitro. Furthermore, we demonstrated that DCLK1 was substan

tially 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.

Conclusions 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 efective therapeutic approach.

摘要：

背景:膽管癌(CCA)具有高度異質(zhì)性和極惡性的特點，預后較差。雙皮質(zhì)素樣激酶1 (DCLK1)促進多種惡性腫瘤的進展。靶向DCLK1或其相關的調(diào)控途徑可以防止幾種惡性腫瘤的發(fā)生和惡化。然而，DCLK1在CCA進展中的作用及其分子機制尚不清楚。因此，我們旨在研究DCLK1是否以及如何促進CCA進展。

方法:采用免疫組化(IHC)法檢測CCA患者DCLK1的表達。我們建立DCLK1敲除細胞系和DCLK1過表達細胞系，進行集落形成實驗和Transwell實驗，探討DCLK1的促腫瘤作用。采用RT-PCR、Western blot和多重熒光染色法評估DCLK1與上皮-間質(zhì)轉(zhuǎn)化(EMT)標志物的相關性。進行RNA測序和生物信息學分析，以確定DCLK1調(diào)節(jié)CCA進展和EMT程序的潛在機制。

結果:DCLK1在CCA組織中過表達，與不良預后相關。DCLK1過表達促進了CCA細胞的侵襲、遷移和增殖，而DCLK1敲低逆轉(zhuǎn)了CCA細胞的惡性傾向，這在體內(nèi)和體外都得到了證實。此外，我們證明DCLK1與EMT計劃的進展有實質(zhì)性的聯(lián)系，其中包括間充質(zhì)標記物的過表達和上皮標記物的下調(diào)。對于潛在的機制，我們提出PI3K/AKT/mTOR通路是DCLK1在腫瘤進展和EMT程序發(fā)生中的關鍵過程。當使用PI3K/AKT/mTOR通路抑制劑LY294002時，腫瘤的增殖、遷移和侵襲能力被極大地抑制，EMT過程通常被逆轉(zhuǎn)。

結論:DCLK1通過PI3K/AKT/mTOR通路促進CCA細胞的惡性生物學行為。在高水平表達DCLK1的膽管癌患者中，抑制PI3K/AKT/mTOR信號通路可能是一種有效的治療方法。

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