Forskolin promotes vasculogenic mimicry and invasion via Notch-1-activated epithelial-to-mesenchymal transition in syncytiolization of trophoblast cells in choriocarcinoma
Forskolin promotes vasculogenic mimicry and invasion via Notch-1-activated epithelial-to-mesenchymal transition in syncytiolization of trophoblast cells in choriocarcinoma
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DOI：
10.3892/ijo.2020.4997
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发表时间：
2020-05-01
影响因子：
5.2
通讯作者：
An, Ruifang
中科院分区：
文献类型：
--
作者：
Xue, Yan;Sun, Rong;An, Ruifang
绒毛膜癌(CC)与其他女性生殖器肿瘤相比，具有较早的血行转移和较高的大出血发生率。血管生成拟态与肿瘤的转移密切相关，合体滋养细胞参与了结直肠癌血管生成的形成。Forsklin是cAMP途径的典型激活剂，参与滋养层细胞的合胞化。为探讨Forsklin在体内外合胞化过程中对JEG-3和JAR细胞侵袭和侵袭的影响及其机制，用100muM Forsklin处理JEG-3和JAR细胞48h，通过伤口愈合和侵袭实验验证细胞的迁移和侵袭能力。建立了3D培养和管状形成试验来检测VM。观察上皮向间充质转化(EMT)的形态和标志物的变化，并研究Forsklin处理的CC细胞中Notch信号通路的作用。本研究结果表明，100亩M forsklin可诱导滋养层细胞合胞化，并增强JEG-3和JAR细胞系的迁移和侵袭能力。此外，与各自的对照组相比，Forskolin在体外和体内均能显著增加Vm的能力，而降低Vm的成管能力。在合胞化过程中，细胞形态呈现EMT改变，包括E-钙粘蛋白和细胞角蛋白的表达下调，N-钙粘蛋白、波形蛋白和锌指E-box结合同源盒1的上调。在Forsklin诱导的CC细胞的VM过程中，Notch-1信号通路被激活以诱导EMT，而用伽玛分泌酶抑制剂DAPT阻断Notch-1通路可以逆转VM和EMT。总体而言，本研究的结果表明，Forsklin通过Notch-1激活的EMT在滋养层细胞的合胞化过程中增强了VM的形成和转移能力。
Choriocarcinoma (CC) is characterized by earlier blood metastasis compared with other female genital tumors and a high incidence of massive hemorrhage. Vasculogenic mimicry (VM) is highly associated with metastasis, and syncytiotrophoblast is involved in the formation of VM in CC. Forskolin is a typical activator of the cAMP pathway, which is involved in the syncytiolization of trophoblastic cells. In the present study, to determine the effects and mechanism of forskolin on cell invasion and VM during syncytiolization in vitro and in vivo, JEG-3 and JAR cell lines were treated with 100 mu M forskolin for 48 h, and wound healing and invasion assays were used to verify cell migratory and invasive capacities. A 3D culture and tube formation assays were established to detect VM. Variation of morphology and markers of the epithelial-to-mesenchymal transition (EMT) were assessed, and the role of the Notch signaling pathway was investigated in CC cells treated with forskolin. The results of the present study demonstrated that 100 mu M forskolin induced syncytiolization of trophoblastic cells and enhanced the migratory and invasive abilities of JEG-3 and JAR cell lines. In addition, the capacity of VM was significantly increased, whereas tube formation ability was decreased by forskolin in vitro and in vivo compared with the respective control groups. The cellular morphology exhibited EMT during the syncytiolization process, which was further supported by the changes in EMT marker expression, including downregulation of E-cadherin and cytokeratin and upregulation of N-cadherin, vimentin and zinc finger E-box-binding homeobox 1. The Notch-1 signaling pathway was activated to induce EMT in forskolin-induced VM process in CC cells, and VM and EMT could be reversed by using the gamma-secretase inhibitor DAPT to block the Notch-1 pathway. Overall, the results of the present study demonstrated that forskolin enhanced the capacity of VM formation and metastasis through Notch-1-activated EMT in the syncytiolization of trophoblastic cells.