Abstract
Verticillin A is a diketopiperazine compound which was previously isolated from Amanita flavorubescens Alk (containing parasitic fungi Hypomyces hyalines (Schw.) Tul.). Here, we initially found, by wound healing assay and Transwell assay in vitro, that verticillin A possesses an inhibitory effect against the migration and invasion of the human colon cancer cell. Subsequently, c-mesenchymal-epithelial transition factor (c-Met) was identified as a molecular target of verticillin A by screening key genes related to cell migration. Verticillin A-mediated c-Met suppression is at the transcriptional level. Further study demonstrated that verticillin A suppressed c-MET phosphorylation and decreased c-MET protein level. In addition, verticillin A inhibited the phosphorylation of c-MET downstream molecules including rat sarcoma (Ras)-associated factor (Raf), extracellular signal-regulated kinase (ERK), and protein kinase B (AKT). Overexpression of Erk partially reversed the verticillin A-mediated anti-metastasis action in the human colon cancer cell. More importantly, verticillin A also inhibited cancer cell metastasis in vivo. Thus, verticillin A can significantly inhibit the migration and invasion of colon cancer cells by targeting c-Met and inhibiting Ras/Raf/mitogen-activated extracellular signal-regulated kinase (MEK)/ERK signaling pathways. Therefore, we determined that verticillin A is a natural compound that can be further developed as an anti-metastatic drug in human cancers.
摘要
目的
结肠癌是全球第三大常见癌症,转移是结肠癌患 者死亡的主要原因,因此迫切需要寻找能够抑制 结肠癌转移新方法。在前期研究中,我们证明了 轮枝孢菌素A 在各种类型的癌症中均具有强大 的抗肿瘤增殖作用。在本研究中,我们试图确定 轮枝孢菌素A 对结肠癌细胞的抗转移作用,并阐 明其潜在的分子机制.
创新点
首次发现轮枝孢菌素A 可以有效地抑制结肠癌细 胞在体内外的迁移和侵袭。在分子水平上,沉默 c-Met 显著降低了结肠细胞的侵袭,而轮枝孢菌 素A 正是有效的c-Met 抑制剂; 同时,轮枝孢菌 素A 还可以通过削弱c-MET 磷酸化来抑制 c-MET 下游Ras/Raf/MEK/ERK 信号通路,从而 达到抗肿瘤转移效果.
方法
采用划痕分析和Transwell 法来证明轮枝孢菌素A 对结肠癌细胞体外迁移和侵袭的影响; 采用蛋白 质印迹法来确定c-MET 及其下游分子的表达水 平和活性; 采用实时定量聚合酶链式反应 (qRT-PCR)分析以评估c-Met 的mRNA 表达水 平; 采用RNA 干扰测定用于沉默细胞中c-Met 的表达; 采用质粒瞬时转染以过度表达Erk.
结论
轮枝孢菌素A 显著抑制结肠癌细胞的迁移和侵 袭,抑制c-Met 转录表达,削弱c-MET 磷酸化, 从而抑制其下游Ras/Raf/MEK/ERK 信号通路, 这是轮枝孢菌素A 抗结肠癌细胞转移的主要分 子机制。根据本研究结果,我们确定天然产物轮 枝孢菌素A 具有进一步开发抗癌症转移的巨大 潜力.
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Change history
16 April 2022
An Erratum to this paper has been published: https://doi.org/10.1631/jzus.B20e0190
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Fei-yan LIU designed study and analyzed data; Fei-yan LIU and Jing-xin LU wrote the manuscript; Qian-qian LIU and Xue-li ZENG performed experiments; Yue-lin GUAN performed plasmid transient transfection; Kai TU provided in vivo study. All authors have read and approved the final version of the manuscript, and, therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Qian-qian LIU, Xue-li ZENG, Yue-lin GUAN, Jing-xin LU, Kai TU, and Fei-yan LIU declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed.
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Liu, Qq., Zeng, Xl., Guan, Yl. et al. Verticillin A inhibits colon cancer cell migration and invasion by targeting c-Met. J. Zhejiang Univ. Sci. B 21, 779–795 (2020). https://doi.org/10.1631/jzus.B2000190
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DOI: https://doi.org/10.1631/jzus.B2000190