School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, China.
College of Pharmacy, Hubei University of Chinese Medicine, Wuhan, China.
PLoS One. 2024 Nov 7;19(11):e0310530. doi: 10.1371/journal.pone.0310530. eCollection 2024.
Isoscopoletin is one of the primary metabolites of natural product scoparone, which was reported to against tumor proliferation. The aim of this study was to explore the mechanism of isoscopoletin against hepatocellular carcinoma (HCC).
Transcriptomics was used to reveal the possible pathways of isoscopoletin against HCC in vitro. The potential targets of isoscopoletin against HCC through affecting glycolysis were analyzed by network pharmacology, then the potential binding abilities of isoscopoletin to glycolysis-related proteins were initially verified by high throughput virtual molecular docking. The affinities of isoscopoletin for glycolysis-related proteins were assayed using microscale thermophoresis (MST), which was reverse-validated by inhibiting the binding ability of isoscopoletin to GPD2. Glucose consumption and lactate production were examined to evaluate the effects of isoscopoletin on intracellular glycolysis, and the regulation of glycolysis-related targets by isoscopoletin was detected using RT-qPCR and ELISA kits.
The results of transcriptomics showed that the differentially expressed genes (DEGs) were mainly enriched in glycolysis and other metabolic-related pathways. Network pharmacology and molecular docking revealed that GPD2, GPI, HSP90AA1 and PGK2 were the core targets in the glycolysis process of isoscopoletin against HCC. MST results showed that there was a strong affinity between isoscopoletin and GPD2, GPI, Hsp90α and PGK2. In vitro results showed that isoscopoletin inhibited glucose consumption and lactate production, while regulating the levels of glycolysis-related proteins.
This study suggests that isoscopoletin may exist an anti-tumor effect by regulating the glycolysis-related proteins GPD2, GPI, Hsp90α and PGK2, inhibiting the glycolysis process in HCC cells, then blocking the energy supply of tumor cells.
异虎耳草素是天然产物虎耳草素的主要代谢物之一，据报道其具有抗肿瘤增殖作用。本研究旨在探讨异虎耳草素抗肝癌（HCC）的作用机制。
采用转录组学技术揭示异虎耳草素在体外抗 HCC 的可能途径。通过网络药理学分析异虎耳草素通过影响糖酵解作用于 HCC 的潜在靶点，然后通过高通量虚拟分子对接初步验证异虎耳草素与糖酵解相关蛋白的潜在结合能力。采用微量热泳动（MST）法测定异虎耳草素与糖酵解相关蛋白的亲和力，并用抑制异虎耳草素与 GPD2 结合能力的方法对其进行反向验证。通过检测葡萄糖消耗和乳酸生成来评估异虎耳草素对细胞内糖酵解的影响，并用 RT-qPCR 和 ELISA 试剂盒检测异虎耳草素对糖酵解相关靶标的调节作用。
转录组学结果表明，差异表达基因（DEGs）主要富集在糖酵解和其他代谢相关途径中。网络药理学和分子对接显示，GPD2、GPI、HSP90AA1 和 PGK2 是异虎耳草素抗 HCC 糖酵解过程中的核心靶点。MST 结果表明，异虎耳草素与 GPD2、GPI、Hsp90α 和 PGK2 之间存在较强的亲和力。体外实验结果表明，异虎耳草素抑制葡萄糖消耗和乳酸生成，同时调节糖酵解相关蛋白水平。
本研究表明，异虎耳草素可能通过调节糖酵解相关蛋白 GPD2、GPI、Hsp90α 和 PGK2 发挥抗肿瘤作用，抑制 HCC 细胞的糖酵解过程，从而阻断肿瘤细胞的能量供应。
