目的:越来越多的证据表明FGFR2是包括胆管癌(CCA)在内的癌症的有吸引力的治疗靶点。本研究调查了FGF10配体激活CCA细胞中FGFR2的致癌机制,并确定了FGFR抑制剂是否可以抑制FGF10介导的CCA细胞迁移。
方法:FGF10对细胞增殖的影响,迁移,使用克隆和transwell测定评估KKU-M213A细胞的侵袭。通过免疫印迹和抗体阵列分析确定FGFR2和促血管生成因子的蛋白表达水平。使用小干扰RNA的FGFR2敲低用于验证FGF10在通过FGFR2促进细胞迁移中的作用。恩替吉替尼(FGFR抑制剂)对细胞活力的影响,在KKU-100,KKU-M213A中测定,KKU-452细胞。此外,在KKU-M213A细胞中评估了FGFR抑制剂通过FGF10/FGFR2刺激抑制迁移的功效.
结果:FGF10能显著增加FGFR/FGFR2的表达,促进细胞增殖,迁移,和KKU-M213A细胞的侵袭。FGF10增加了p-Akt的表达水平,p-mTOR,VEGF,子弹,和与转移相关的促血管生成蛋白。由FGF10介导的细胞迁移在FGFR2敲低细胞中显著降低。此外,FGF10/FGFR2促进细胞迁移,被FGFR抑制剂抑制。
结论:FGF10/FGFR2激活Akt/mTOR和VEGF/Slug通路,与CCA中迁移和入侵的刺激有关。此外,FGFR抑制剂抑制了FGF10/FGFR2信号传导,从而抑制了细胞迁移,这需要进一步研究它们在CCA治疗中的临床应用。
OBJECTIVE: Increasing evidence has revealed FGFR2 as an attractive therapeutic target for cancer including cholangiocarcinoma (CCA). The present study investigated the oncogenic mechanisms by which
FGF10 ligand activates FGFR2 in CCA cells and determined whether FGFR inhibitors could suppress
FGF10-mediated migration of CCA cells.
METHODS: Effects of
FGF10 on the proliferation, migration, and invasion of KKU-M213A cells were assessed using clonogenic and transwell assays. Protein expression levels of FGFR2 and pro-angiogenic factors were determined via immunoblotting and antibody array analysis. FGFR2 knockdown using a small interfering RNA was used to validate the role of FGF10 in promoting cell migration via FGFR2. The effects of infigratinib (FGFR inhibitor) on cell viability, were determined in KKU-100, KKU-M213A, KKU-452 cells. Moreover, the efficacy of the FGFR inhibitor in suppressing migration via FGF10/FGFR2 stimulation was assessed in KKU-M213A cells.
RESULTS: FGF10 significantly increased the expression of phospho-FGFR/FGFR2 and promoted the proliferation, migration, and invasion of KKU-M213A cells. FGF10 increased the expression levels of p-Akt, p-mTOR, VEGF, Slug, and pro-angiogenic proteins related to metastasis. Cell migration mediated by
FGF10 was markedly decreased in FGFR2-knockdown cells. Moreover,
FGF10/FGFR2 promoted the migration of cells, which was suppressed by the FGFR inhibitor.
CONCLUSIONS: FGF10/FGFR2 activates the Akt/mTOR and VEGF/Slug pathways, which are associated with the stimulation of migration and invasion in CCA. Moreover, the FGF10/FGFR2 signaling was inhibited by an FGFR inhibitor resulting suppression of cell migration, which warrants further studies on their clinical utility for CCA treatment.