Lyu Shao-Cheng, Han Dong-Dong, Li Xian-Liang, Ma Jun, Wu Qiao, Dong Hong-Meng, Bai Chun, He Qiang
Department of Hepatobiliary Surgery, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, P.R. China.
Oncol Lett. 2018 Feb;15(2):2085-2090. doi: 10.3892/ol.2017.7542. Epub 2017 Dec 7.
Cholangiocarcinoma (CCA) is a rare and fatal tumor. In previous decades, there has been a steady increase in the incidence and mortality rates of this tumor worldwide. Metastasis is regarded as the major factor that contributes to poor prognosis in CCA patients. Studies therefore aim to develop novel therapeutic targets to control CCA metastasis. Fyn is known to enhance expression and promote metastasis in various cancers, including pancreatic cancer, prostate cancer and colorectal cancer. However, the exact function and mechanism of Fyn in CCA metastasis remains unclear. In the present study, mRNA and protein expression levels of Fyn, AMP-activated protein kinase (AMPK), phosphorylated (p-)AMPK, mammalian target of rapamycin (mTOR) and p-mTOR were measured, using the reverse transcription-quantitative polymerase chain reaction and western blot analysis, in CCA tissues and cell lines. In addition, Transwell assays were used to determine the migratory and invasive abilities of human CCA QBC939, following transfection. In the present study, it was found that Fyn was overexpressed in CCA cell lines. Fyn knockdown inhibited CCA cell migration and invasion. Furthermore, it was demonstrated that Fyn knockdown induces phosphorylation of AMPK, inhibits downstream phosphorylation of mTOR, and activate the AMPK/mTOR signaling pathway. Compound C, an AMPK inhibitor, inhibited the AMPK/mTOR signaling pathway, and reversed the effect of Fyn knockdown on migration and invasion of CCA cells. In conclusion, the present study suggests that Fyn knockdown inhibits cell migration and invasion by regulating the AMPK/mTOR signaling pathway in CCA cell lines and that Fyn knockdown is a potential target for anti-CCA therapy.
胆管癌（CCA）是一种罕见的致命肿瘤。在过去几十年中，全球范围内该肿瘤的发病率和死亡率一直在稳步上升。转移被认为是导致CCA患者预后不良的主要因素。因此，研究旨在开发新的治疗靶点来控制CCA转移。已知Fyn可增强多种癌症（包括胰腺癌、前列腺癌和结直肠癌）中的表达并促进转移。然而，Fyn在CCA转移中的确切功能和机制仍不清楚。在本研究中，使用逆转录定量聚合酶链反应和蛋白质印迹分析，测量了CCA组织和细胞系中Fyn、AMP激活的蛋白激酶（AMPK）、磷酸化（p-）AMPK、雷帕霉素靶蛋白（mTOR）和p-mTOR的mRNA和蛋白表达水平。此外，采用Transwell实验来确定转染后人CCA QBC939细胞的迁移和侵袭能力。在本研究中，发现Fyn在CCA细胞系中过表达。敲低Fyn可抑制CCA细胞的迁移和侵袭。此外，还证明敲低Fyn可诱导AMPK磷酸化，抑制mTOR的下游磷酸化，并激活AMPK/mTOR信号通路。AMPK抑制剂Compound C可抑制AMPK/mTOR信号通路，并逆转敲低Fyn对CCA细胞迁移和侵袭的影响。总之，本研究表明敲低Fyn通过调节CCA细胞系中的AMPK/mTOR信号通路来抑制细胞迁移和侵袭，并且敲低Fyn是抗CCA治疗的一个潜在靶点。
