Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Biochemistry and Molecular Biology, Peking University Cancer Hospital & Institute, Beijing, 100142, China.
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
Acta Pharmacol Sin. 2023 Jan;44(1):189-200. doi: 10.1038/s41401-022-00940-4. Epub 2022 Jul 1.
The high incidence of lymphatic metastasis is closely related to poor prognosis and mortality in cancers. Potent inhibitors to prevent pathological lymphangiogenesis and lymphatic spread are urgently needed. The VEGF-C-VEGFR3 pathway plays a vital role in driving lymphangiogenesis and lymph node metastasis. In addition, COX2 in tumor cells and tumor-associated macrophages (TAMs) facilitates lymphangiogenesis. We recently reported that aiphanol, a natural stilbenolignan, attenuates tumor angiogenesis by repressing VEGFR2 and COX2. In this study, we evaluated the antilymphangiogenic and antimetastatic potency of aiphanol using in vitro, ex vivo and in vivo systems. We first demonstrated that aiphanol directly bound to VEGFR3 and blocked its kinase activity with an half-maximal inhibitory concentration (IC) value of 0.29 μM in an in vitro ADP-Glo kinase assay. Furthermore, we showed that aiphanol (7.5-30 μM) dose-dependently counteracted VEGF-C-induced proliferation, migration and tubular formation of lymphatic endothelial cells (LECs), which was further verified in vivo. VEGFR3 knockdown markedly mitigated the inhibitory potency of aiphanol on lymphangiogenesis. In 4T1-luc breast tumor-bearing mice, oral administration of aiphanol (5 and 30 mg· kg ·d) dose-dependently decreased lymphatic metastasis and prolonged survival time, which was associated with impaired lymphangiogenesis, angiogenesis and, interestingly, macrophage infiltration. In addition, we found that aiphanol decreased the COX2-dependent secretion of PGE2 and VEGF-C from tumor cells and macrophages. These results demonstrate that aiphanol is an appealing agent for preventing lymphangiogenesis and lymphatic dissemination by synergistically targeting VEGFR3 and inhibiting the COX2-PGE2-VEGF-C signaling axis.
淋巴转移的高发生率与癌症的不良预后和死亡率密切相关。迫切需要有效的抑制剂来预防病理性淋巴管生成和淋巴扩散。VEGF-C-VEGFR3 通路在驱动淋巴管生成和淋巴结转移中起着至关重要的作用。此外，肿瘤细胞和肿瘤相关巨噬细胞（TAMs）中的 COX2 促进淋巴管生成。我们最近报道，天然二苯乙烯类化合物 aiphanol 通过抑制 VEGFR2 和 COX2 来减弱肿瘤血管生成。在这项研究中，我们使用体外、离体和体内系统评估了 aiphanol 的抗淋巴管生成和抗转移作用。我们首先证明 aiphanol 可直接与 VEGFR3 结合，并在体外 ADP-Glo 激酶测定中以半最大抑制浓度（IC）值 0.29μM 阻断其激酶活性。此外，我们表明 aiphanol（7.5-30μM）剂量依赖性地拮抗 VEGF-C 诱导的淋巴管内皮细胞（LEC）增殖、迁移和管状形成，这在体内得到了进一步验证。VEGFR3 敲低显著减轻了 aiphanol 对淋巴管生成的抑制作用。在 4T1-luc 乳腺癌荷瘤小鼠中，口服给予 aiphanol（5 和 30mg·kg·d）剂量依赖性地减少了淋巴转移并延长了生存时间，这与淋巴管生成、血管生成的受损有关，有趣的是，还与巨噬细胞浸润有关。此外，我们发现 aiphanol 降低了肿瘤细胞和巨噬细胞中 COX2 依赖性 PGE2 和 VEGF-C 的分泌。这些结果表明，aiphanol 通过协同靶向 VEGFR3 和抑制 COX2-PGE2-VEGF-C 信号轴，是一种有吸引力的预防淋巴管生成和淋巴扩散的药物。
