Network Pharmacology and Experimental Evidence Reveal Dioscin Suppresses Proliferation, Invasion, and EMT via AKT/GSK3b/mTOR Signaling in Lung Adenocarcinoma
Network Pharmacology and Experimental Evidence Reveal Dioscin Suppresses Proliferation, Invasion, and EMT via AKT/GSK3b/mTOR Signaling in Lung Adenocarcinoma
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网络药理学和实验证据表明薯蓣皂苷通过 AKT/GSK3b/mTOR 信号传导抑制肺腺癌的增殖、侵袭和 EMT
DOI：
10.2147/dddt.s249651
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发表时间：
2020-01-01
影响因子：
4.8
通讯作者：
He, Yanli
中科院分区：
文献类型：
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作者：
Mao, Wenli;Yin, Heng;He, Yanli
目的：薯蓣皂苷是一种来源于多种植物的天然苷类化合物，具有抗癌活性。几项研究发现，它逆转TGF-β 1诱导的上皮-间充质转化（EMT）。薯蓣皂苷是否可以逆转EMT的途径以外的TGF-β。方法：我们使用基于网络的药理学方法，系统地探讨薯蓣皂苷作用于肺癌的潜在机制。采用Cell Counting Kit-8法、划痕愈合法、Transwell法、Matrigel侵袭法、免疫荧光法和Western blotting法对薯蓣皂苷的关键作用靶点进行了预测，并对薯蓣皂苷对EMT的影响进行了验证。在具有最低p值的20个中，PI 3 K-AKT信号传导通路参与并且与EMT显著相关。在化合物-靶标分析中，AKT 1和mTOR以高程度（反映更高的连接性）参与PI 3 K-AKT信号传导途径。分子对接表明薯蓣皂苷-AKT 1和薯蓣皂苷-mTOR结合的发生。功能实验表明，薯蓣皂苷抑制人肺腺癌细胞的增殖，迁移，侵袭和EMT的剂量依赖性的方式，没有TGF-β刺激。此外，我们确定薯蓣皂苷下调人肺腺癌细胞中的p-AKT、p-mTOR和p-GSK 3 β，而不影响其总蛋白水平。结论：薯蓣皂苷通过抑制AKT/mTOR/GSK 3 β信号通路，抑制肺腺癌细胞增殖、侵袭和EMT，其机制可能是通过与AKT和mTOR结合，抑制其磷酸化。
Purpose: Dioscin, a natural glycoside derived from many plants, has been proved to exert anti-cancer activity. Several studies have found that it reverses TGF-beta 1-induced epithelial-mesenchymal transition (EMT). Whether dioscin can reverse EMT by pathways other than TGF-beta is still unknown.Methods: We used network-based pharmacological methods to systematically explore the potential mechanisms by which dioscin acts on lung cancer. Cell Counting Kit-8 assay, scratch healing, Transwell assay, Matrigel invasion assay, immunofluorescence assay, and Western blotting were employed to confirm the prediction of key targets and the effects of dioscin on EMT.Results: Here, using network-based pharmacological methods, we found 42 possible lung cancer-related targets of dioscin, which were assigned to 98 KEGG pathways. Among the 20 with the lowest p-values, the PI3K-AKT signaling pathway is involved and significantly related to EMT. AKT1 and mTOR, with high degrees (reflecting higher connectivity) in the compound-target analysis, participate in the PI3K-AKT signaling pathway. Molecular docking indicated the occurrence of dioscin-AKT1 and dioscin-mTOR binding. Functional experiments demonstrated that dioscin suppressed the proliferation, migration, invasion, and EMT of human lung adenocarcinoma cells in a dose-dependent manner, without TGF-beta stimulation. Furthermore, we determined that dioscin downregulated p-AKT, p-mTOR and p-GSK3 beta in human lung adenocarcinoma cells without affecting their total protein levels. The PI3K inhibitor LY294002 augmented these changes.Conclusion: Dioscin suppressed proliferation, invasion and EMT of lung adenocarcinoma cells via the inactivation of AKT/mTOR/GSK3 beta signaling, probably by binding to AKT and mTOR, and inhibiting their phosphorylation.