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Exploring the effect of Huangqi Fuling Decoction on gastric cancer based on UPLC-MS, network pharmacology and
Doudou Lu1, Ling Yuan2, Guoqing Chen2
1School of Basic Medical Sciences, Ningxia Medical University, Yinchuan, Ningxia, 750004, China.
Background:
Huangqi Fuling Decoction (HQFLT) is a traditional Chinese medicine prescription that is applied to treat a variety of diseases. However, the mechanism of HQFLT in the treatment of gastric cancer is still unclear, which hinders its clinical application.
Methods:
Components and targets of HQFLT were collected from TCMSP, TCMID and ETCM databases, and potential targets of gastric cancer were collected from GEO database. The intersection targets of component disease were imported into STRING database to construct protein-protein interaction network, and the intersection targets were analyzed by GO and KEGG to explore the involved biological processes. The network topology analysis of intersection targets was carried out using Cytoscape software, and hub genes were selected according to the degree value. To further identify the main active components of HQFLT, we used UPLC-MS/MS techniques for analysis and intersected with network pharmacological results to obtain the most critical components. Molecular docking techniques were used to calculate the binding energy of intersection components and hub genes. Regarding cell experiments, the effect of HQFLT on the viability of gastric cancer cells was detected by CCK-8 assay, and the effect of HQFLT on cell proliferation was verified by flow cytometry and colony formation assay. The transwell experiment was conducted to investigate the effect of HQFLT on tumor metastasis. qRT-PCR and Western blot were used to detect the regulatory effects of HQFLT on hub genes and key signaling pathways.
Results:
A total of 189 active components, and 278 targets of HQFLT and 1817 targets of gastric cancer were obtained by network pharmacology. A total of 51 intersection targets were screened by Venny method. These targets were mainly involved in signal transduction, apoptosis regulation and angiogenesis, and may be involved in the regulation of MAPK signaling pathway. A total of 5 hub genes were obtained after degree screening by Cytoscape, namely IL6, PTGS2, ICAM1, VCAMI and CCL2, which showed significant differences in clinical staging, molecular typing and survival prognosis. After the intersection of 44 components obtained from UPLC-MS and network pharmacology, Calycosin was obtained. The results of molecular docking showed that the compound had strong binding ability with hub genes. Regarding cell experiments, HQFLT had the ability to inhibit proliferation, block cell cycle, induce cell apoptosis, inhibit colony formation, and inhibit migration and invasion of gastric cancer cells. qRT-PCR and Western blot experiments confirmed that HQFLT could down-regulate the expression of hub genes and inhibit the activation of RAS-MER-ERK signaling pathway.
Conclusion:
HQFLT inhibited the proliferation and metastasis of gastric cancer. The mechanism may involve the regulation of the RAS-MEK-ERK signaling pathway, with ICAM1 and VCAM1 being potential targets for targeted therapy.
