Chen Hsien-Lin, Su Yu-Chieh, Chen Huang-Chi, Su Jui-Hsin, Wu Chang-Yi, Wang Shih-Wei, Lin In-Pin, Chen Chung-Yi, Lee Chien-Hsing
Division of General Surgery, Department of Surgery, Chi-Mei Medical Center, Liouying, Tainan 73657, Taiwan.
Department of Medicine, School of Medicine, I-Shou University, Kaohsiung 840203, Taiwan.
Biomedicines. 2021 Aug 29;9(9):1109. doi: 10.3390/biomedicines9091109.
Lymphatic metastasis is a biological procedure associated with the pathogenesis of several diseases, especially in tumor metastasis. Therefore, regulation of lymphangiogenesis has become a promising strategy for cancer therapy. In this study, we aimed to investigate the anti-lymphangiogenic effect of heteronemin (SP-1) isolated from the sponge sp. in vitro and in vivo. Human lymphatic endothelial cells (LECs) were utilized to evaluate the anti-lymphangiogenic effect of SP-1 in vitro. Molecular docking, western blotting, flow-cytometry, MTT and ELISA were performed to investigate the mechanism of action. For in vivo approaches, the transgenic (fli1:EGFP; gata1:DsRed) zebrafish and mouse ear sponges were used. Molecular docking studies showed that SP-1 is a potent vascular endothelial growth factor receptor 3 (VEGFR-3)-binding compound. Treatment of LEC with SP-1 reduced the phosphorylation of VEGFR-3. SP-1 suppressed the development of the thoracic duct in zebrafish and mouse lymphangiogenesis ear sponges in vivo. Mechanistically, SP-1 induced the cell cycle arrest of LECs in the G0/G1 phase and reduced the downstream of VEGFR-3, such as phosphorylated MEK/ERK and NF-κB. In addition, SP-1 inhibited LECs' tubulogenesis and migration through the ARF-1 and MMP-9/VE-cadherin/vimentin. Overall, anti-lymphangiogenic properties of SP-1 occur by downregulating the VEGFR-3 cascade, ARF-1 and MMP-9/VE-cadherin/vimentin. Collectively, these results proposed that SP-1 might be a potential candidate for the treatment of lymphangiogenesis-associated diseases.
淋巴转移是一种与多种疾病发病机制相关的生物学过程，尤其是在肿瘤转移方面。因此，调控淋巴管生成已成为一种有前景的癌症治疗策略。在本研究中，我们旨在研究从海绵sp.中分离出的异壬素（SP-1）在体外和体内的抗淋巴管生成作用。利用人淋巴管内皮细胞（LECs）评估SP-1在体外的抗淋巴管生成作用。进行分子对接、蛋白质免疫印迹、流式细胞术、MTT和酶联免疫吸附测定以研究其作用机制。对于体内实验方法，使用转基因（fli1:EGFP；gata1:DsRed）斑马鱼和小鼠耳海绵。分子对接研究表明，SP-1是一种有效的血管内皮生长因子受体3（VEGFR-3）结合化合物。用SP-1处理LEC可降低VEGFR-3的磷酸化水平。SP-1在体内抑制斑马鱼胸导管的发育以及小鼠淋巴管生成耳海绵中的淋巴管生成。从机制上讲，SP-1诱导LECs细胞周期停滞在G0/G1期，并减少VEGFR-3的下游分子，如磷酸化的MEK/ERK和NF-κB。此外，SP-1通过ARF-1和MMP-9/VE-钙黏蛋白/波形蛋白抑制LECs的管腔形成和迁移。总体而言，SP-1的抗淋巴管生成特性是通过下调VEGFR-3级联反应、ARF-1和MMP-9/VE-钙黏蛋白/波形蛋白实现的。综上所述，这些结果表明SP-1可能是治疗淋巴管生成相关疾病的潜在候选药物。
