Integrating UPLC-HR-MS/MS, Network Pharmacology, and Experimental Validation to Uncover the Mechanisms of Jin’gan Capsules against Breast Cancer

Abstract

In the theory of traditional Chinese medicine (TCM), “liver-qi” stagnation and heat-induced toxicity represent the main etiologies of breast cancer. Recently, several TCMs with heat-clearing and detoxification efficacy have shown inhibitory effects on breast cancer. Jin’gan capsules (JGCs), initially approved to treat colds in China, are a heat-clearing and detoxification TCM formula. However, the anticancer activity of JGCs against breast cancer and its underlying mechanisms remain unclear. First, we assessed the antiproliferative activity of JGCs in breast cancer cell lines and evaluated their effects on cell apoptosis and the cell cycle by flow cytometry. Furthermore, we identified the potential bioactive components of JGCs and their corresponding target genes and constructed a bioactive compound–target interaction network by ultra-performance liquid chromatography–high-resolution tandem mass spectrometry (UPLC-HR-MS/MS) and network pharmacology analysis. Finally, the underlying mechanism was investigated through gene function enrichment analysis and experimental validation. We found that JGCs significantly inhibited breast cancer cell growth with IC 50 values of 0.56 ± 0.03, 0.16 ± 0.03, and 0.94 ± 0.09 mg/mL for MDA-MB-231, MDA-MB-468, and MCF-7, respectively. In addition, JGC treatment dramatically induced apoptosis and S phase cell cycle arrest in breast cancer cells. Western blot analysis confirmed that JGCs could regulate the protein levels of apoptosis- and cell cycle-related genes. Utilizing UPLC-HR-MS/MS analysis and network pharmacology, we identified 7 potential bioactive ingredients in JGCs and 116 antibreast cancer targets. Functional enrichment analysis indicated that the antitumor effects of JGCs were strongly associated with apoptosis and the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway. Western blot analysis validated that JGC treatment markedly decreased the expression levels of p -JAK2, p -STAT3, and STAT3. Our findings suggest that JGCs suppress breast cancer cell proliferation and induce cell cycle arrest and apoptosis partly by inhibiting the JAK2/STAT3 signaling pathway, highlighting JGCs as a potential therapeutic candidate against breast cancer.

Fang-Xia-Dihuang decoction inhibits breast cancer progression induced by psychological stress via down-regulation of PI3K/AKT and JAK2/STAT3 pathways: An in vivo and a network pharmacology assessment

“…However, FXDH inhibited the overactivation of the PI3K/AKT and JAK2/STAT3 pathways. This finding supports the growing evidence indicating that TCM molecules can inhibit tumor growth by downregulating the PI3K/AKT and JAK2/STAT pathways (Zhang et al, 2019;Xu et al, 2020;Qiu et al, 2022). Remarkably, FXDH had a regulatory effect on depressionlike behavior in mice.…”

Exploring the effect and mechanism of DaYuan Yin against acute lung injury by Network Pharmacology,molecular docking and experiment validation

“…GO and KEGG pathway enrichment analysis of the main targets of DYY and ALI were performed using Metascape(https://metascape.org/) [29]. Based on the P-value less than 0.05,the top 20 of biological process (BP), cellular component (CC), molecular function (MF) terms and the top 30 of KEGG terms were imported and visualized on the bioinformatics platform (http://www.bioinformatics.com.cn/) to analyze the key molecular biology processes and key targets' signal path.…”

Traditional Chinese Medicine for Breast Cancer: A Review

“… 70 Jin’gan capsules (JGCs) suppress BC cell proliferation and induce cell cycle arrest and apoptosis partly by inhibiting the JAK2/STAT3 signaling pathway. 77 The effective ingredients of Xiao ai Jie du decoction: quercetin and ursolic acid, through the combination of both, the mitochondrial membrane potential is notably reduced and the activity of PI3K Akt signaling pathway is inhibited, which induces cancer cell death. 25 Taohong Siwu Decoction (TSD) can significantly inhibit KRAS signaling in cancer and activation of the apoptosis pathway, p53 pathway, and hypoxia pathways inhibits the growth and spread of cancer cells, induction of G1 arrest and apoptosis of MDA-MB-231 cells can regulate the expression of MYC, BIRC5, EGF and PIK3R1.…”

“…Alkali ethanol 67 Baicalein, 68 Goushecao, 69 Guizhi Fuling Decoction, 70 Xiao ai Jie du decoction 25 PI3K/AKT relevant pathway Tengligen 71 AKT/GSK-3b pathway Ai du Qing formula 72 TAM/CXCL1/Treg pathway Berberine 73 Ephrin-B2 signaling pathway Xihuang Pill, 74,75 Ethyl gallate 76 NF-κB signaling pathway Jin'gan capsules, 77…”