Dai Yifei, Qiang Weijie, Yu Xiankuo, Cai Siwei, Lin Kequan, Xie Lan, Lan Xun, Wang Dong
Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing 100084, China.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China.
Comput Struct Biotechnol J. 2020 May 18;18:1121-1136. doi: 10.1016/j.csbj.2020.05.004. eCollection 2020.
As one of the classical traditional Chinese medicine (TCM) prescriptions in treating gynecological tumors, Guizhi Fuling Decoction (GFD) has been used to treat breast cancer (BRCA). Nonetheless, the potential molecular mechanism remains unclear so far. Therefore, systems pharmacology was used in combination with high throughput sequencing-based high throughput screening (HTS) assay and bioinformatic technologies in this study to investigate the molecular mechanisms of GFD in treating BRCA. By computationally analyzing 76 active ingredients in GFD, 38 potential therapeutic targets were predicted and significantly enriched in the "pathways in cancer". Meanwhile, experimental analysis was carried out to examine changes in the expression levels of 308 genes involved in the "pathways in cancer" in BRCA cells treated by five herbs of GFD utilizing HTS platform, and 5 key therapeutic targets, including HRAS, EGFR, PTK2, SOS1, and ITGB1, were identified. The binding mode of active compounds to these five targets was analyzed by molecular docking and molecular dynamics simulation. It was found after integrating the computational and experimental data that, GFD possessed the anti-proliferation, pro-apoptosis, and anti-angiogenesis activities mainly through regulating the PI3K and the MAPK signaling pathways to inhibit BRCA. Besides, consistent with the TCM theory about the synergy of (Guizhi) by (Mudanpi) in GFD, both of these two herbs acted on the same targets and pathways. Taken together, the combined application of computational systems pharmacology techniques and experimental HTS platform provides a practical research strategy to investigate the functional and biological mechanisms of the complicated TCM prescriptions.
作为治疗妇科肿瘤的经典中药方剂之一，桂枝茯苓汤已被用于治疗乳腺癌。然而，其潜在的分子机制至今仍不清楚。因此，本研究采用系统药理学结合基于高通量测序的高通量筛选（HTS）分析和生物信息学技术，以探究桂枝茯苓汤治疗乳腺癌的分子机制。通过对桂枝茯苓汤中76种活性成分进行计算分析，预测出38个潜在治疗靶点，并在“癌症通路”中显著富集。同时，利用HTS平台对桂枝茯苓汤五味药处理的乳腺癌细胞中参与“癌症通路”的308个基因的表达水平变化进行实验分析，确定了5个关键治疗靶点，包括HRAS、EGFR、PTK2、SOS1和ITGB1。通过分子对接和分子动力学模拟分析了活性化合物与这5个靶点的结合模式。综合计算和实验数据发现，桂枝茯苓汤主要通过调节PI3K和MAPK信号通路抑制乳腺癌，从而具有抗增殖、促凋亡和抗血管生成活性。此外，与中医关于桂枝茯苓汤中桂枝与牡丹皮协同作用的理论一致，这两味药作用于相同的靶点和通路。综上所述，计算系统药理学技术与实验HTS平台的联合应用为研究复杂中药方剂的功能和生物学机制提供了一种切实可行的研究策略。
