Department of Intervention Hunan, Cancer Hospital, The Affiliated Cancer Hospital of Xiangya School of Medicine Central South University, Changsha, China.
Infection Controlling Center, The Third Xiangya Hospital of Central South University, Changsha, China.
Pharmacology. 2021;106(9-10):509-519. doi: 10.1159/000514410. Epub 2021 Aug 19.
Intrahepatic cholangiocarcinoma (ICC), which is difficult to diagnose and is usually fatal due to its late clinical presentation and a lack of effective treatment, has risen over the past decades but without much improvement in prognosis.
The study aimed to investigate the role of apatinib that targets vascular endothelial growth factor receptor-2 (VEGFR2) in ICC.
MTT assays, cell scratch assays, and tube formation assays were used to assess the effect of apatinib on human ICC cell line (HuCCT-1) and RBE cells proliferation, migration, and angiogenic capacity, respectively. Expression of vascular endothelial growth factor (VEGF), VEGFR2, signal transducer and activator of transcription factor 3 (STAT3), pSTAT3, and hypoxia inducible factor 1 subunit alpha (HIF-1α) pathway proteins was assessed using Western blotting and mRNA expression analysis in HuCCT-1 was performed using RT-qPCR assays. The pcDNA 3.1(-)-VEGFR2 and pcDNA 3.1(-)-HIF-1α were transfected into HuCCT-1 and RBE cells using Lipofectamine 2,000 to obtain overexpressed HuCCT-1 and RBE cells.
We found that apatinib-inhibited proliferation, migration, and angiogenesis of HuCCT-1 and RBE cells in vitro in a dose-dependent manner. We also proved that apatinib effectively inhibits angiogenesis in tumor cells by blocking the expression of VEGF and VEGFR2 in these cells. In addition, we demonstrated that apatinib regulates the expression of STAT3 phosphorylation by inhibiting VEGFR2. Finally, we showed that apatinib regulates ICC angiogenesis and HIF-1α/VEGF expression via STAT3.
Based on the above findings, we conclude that apatinib inhibits HuCCT-1 and RBE cell proliferation, migration, and tumor angiogenesis by inhibiting the VEGFR2/STAT3/HIF-1α axis signaling pathway. Apatinib can be a promising drug for ICC-targeted molecular therapy.
肝内胆管癌（ICC）由于其临床表现较晚且缺乏有效治疗方法，因此难以诊断，通常导致死亡。在过去几十年中，ICC 的发病率有所上升，但预后并未得到明显改善。
本研究旨在探讨靶向血管内皮生长因子受体-2（VEGFR2）的阿帕替尼在 ICC 中的作用。
采用 MTT 检测、细胞划痕实验和管腔形成实验分别评估阿帕替尼对人 ICC 细胞系（HuCCT-1）和 RBE 细胞增殖、迁移和血管生成能力的影响。采用 Western blot 检测血管内皮生长因子（VEGF）、VEGFR2、信号转导和转录激活因子 3（STAT3）、pSTAT3 和缺氧诱导因子 1 亚单位α（HIF-1α）通路蛋白的表达，采用 RT-qPCR 检测 HuCCT-1 中的 mRNA 表达。使用 Lipofectamine 2,000 将 pcDNA 3.1(-)-VEGFR2 和 pcDNA 3.1(-)-HIF-1α 转染至 HuCCT-1 和 RBE 细胞中，获得过表达 HuCCT-1 和 RBE 细胞。
我们发现阿帕替尼以剂量依赖的方式抑制 HuCCT-1 和 RBE 细胞在体外的增殖、迁移和血管生成。我们还证明阿帕替尼通过阻断这些细胞中 VEGF 和 VEGFR2 的表达，有效地抑制肿瘤细胞中的血管生成。此外，我们表明阿帕替尼通过抑制 VEGFR2 调节 STAT3 磷酸化的表达。最后，我们表明阿帕替尼通过 STAT3 调节 ICC 血管生成和 HIF-1α/VEGF 的表达。
基于上述发现，我们得出结论，阿帕替尼通过抑制 VEGFR2/STAT3/HIF-1α 轴信号通路抑制 HuCCT-1 和 RBE 细胞增殖、迁移和肿瘤血管生成。阿帕替尼可能成为一种有前途的 ICC 靶向分子治疗药物。
