Guo Zhenhui, Yin Heng, Wu Tong, Wu Shaofeng, Liu Lingyun, Zhang Lei, He Yanli, Zhang Ren, Liu Na
Department of Medical Biotechnology, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
The First Clinical Medical School, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
J Ethnopharmacol. 2022 Jul 15;293:115280. doi: 10.1016/j.jep.2022.115280. Epub 2022 Apr 8.
Xie Bai San is a Chinese medicine prescription that has been used to treat lung cancer in China for a long time. It has been proven to alleviate the symptoms and extend the survival time of lung cancer patients. Xie Bai San comprises Cortex Lycii, Cortex Mori, and Radix Glycyrrhizae Preparata. The effects and mechanisms of Cortex Mori and Glycyrrhizae on lung cancer have been reported, whereas the underlying mechanism of Cortex Lycii remains unknown.
Network pharmacology was used to explore the unknown mechanisms underlying the effect of Cortex Lycii on lung cancer. Molecular docking was used to predict the binding of a compound to the protein. The fingerprint of Cortex Lycii was obtained by HPLC. Cell counting Kit-8 assay was used to determine the appropriate concentration of Cortex Lycii extract for human lung adenocarcinoma cells, A549 and H1299. Wound healing assay and Matrigel invasion assay were used to detect the influence of Cortex Lycii extract on the migration and invasion ability of A549 and H1299. The protein expression level was detected by western blot and immunohistochemical staining.
Using network pharmacology, 38 components of Cortex Lycii and 79 possible lung cancer-related target genes of Cortex Lycii were obtained. The targets were assigned to 35 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and the PI3K-AKT signaling pathway contained the most targets and had the second-lowest P-value. The molecular docking showed the components of Cortex Lycii bound to HSP90AB1. Among them, 6 components bound to HSP90AB1 in which HSP90AB1 binds to and phosphorylates AKT. The functional experiments showed that Cortex Lycii suppressed the migration and invasion of human lung cancer cells in a dose-dependent manner. Cortex Lycii up-regulated E-Cadherin and down-regulated N-Cadherin, Vimentin, and MMP2. Furthermore, Cortex Lycii made no change in the total AKT and mTOR protein levels, but caused the down-regulation of p-AKT and p-mTOR in human lung cancer cells, which was reversed by Terazosin, an agonist of HSP90. Moreover, acacetin and apigenin, two components of Cortex Lycii, reduced the protein level of p-AKT and p-mTOR, and the reduction was also inhibited by Terazosin.
Cortex Lycii suppressed epithelial-mesenchymal transition (EMT) in lung cancer cells through the PI3K-AKT-mTOR signaling pathway, possibly by targeting HSP90AB1 and inhibiting HSP90AB1-AKT binding.
泻白散是一种在中国长期用于治疗肺癌的中药方剂。已证实其可缓解肺癌患者症状并延长生存期。泻白散由地骨皮、桑白皮和蜜炙甘草组成。桑白皮和甘草对肺癌的作用及机制已有报道，而地骨皮的潜在机制尚不清楚。
采用网络药理学探究地骨皮对肺癌作用的未知机制。利用分子对接预测化合物与蛋白质的结合。通过高效液相色谱法获得地骨皮的指纹图谱。采用细胞计数试剂盒 - 8法确定地骨皮提取物对人肺腺癌细胞A549和H1299的合适浓度。采用伤口愈合试验和基质胶侵袭试验检测地骨皮提取物对A549和H1299迁移和侵袭能力的影响。通过蛋白质印迹法和免疫组织化学染色检测蛋白质表达水平。
利用网络药理学，获得了地骨皮的38种成分和79个可能与肺癌相关的靶基因。这些靶基因被分配到35条京都基因与基因组百科全书（KEGG）通路中，PI3K - AKT信号通路包含的靶基因最多且P值第二低。分子对接显示地骨皮的成分与HSP90AB1结合。其中，6种成分与HSP90AB1结合，HSP90AB1与AKT结合并使其磷酸化。功能实验表明，地骨皮以剂量依赖方式抑制人肺癌细胞的迁移和侵袭。地骨皮上调E - 钙黏蛋白，下调N - 钙黏蛋白、波形蛋白和MMP2。此外，地骨皮对人肺癌细胞中总AKT和mTOR蛋白水平无影响，但导致p - AKT和p - mTOR下调，而HSP90激动剂特拉唑嗪可逆转这种下调。此外，地骨皮的两种成分刺槐素和芹菜素降低了p - AKT和p - mTOR的蛋白水平，且这种降低也被特拉唑嗪抑制。
地骨皮可能通过靶向HSP90AB1并抑制HSP90AB1 - AKT结合，经由PI3K - AKT - mTOR信号通路抑制肺癌细胞的上皮 - 间质转化（EMT）。
