Mechanisms of Gualou Beimu Yin in suppressing the development and metastasis of triple-negative breast cancer: an integrated study based on network pharmacology, transcriptomics, and metabolomics

瓜蒌北木饮抑制三阴性乳腺癌发生发展及转移的机制:基于网络药理学、转录组学和代谢组学的整合研究

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Abstract

OBJECTIVE: To identify the antitumor effects of Gualou Beimu Yin (, GLBMY) in triple-negative breast cancer (TNBC) and to explore the underlying mechanisms. METHODS: A mouse model of breast cancer was established and treated with GLBMY. Freeze-dried GLBMY powder was used to treat MDA-MB-231 and BT549 cells to assess the therapeutic efficacy of GLBMY against TNBC. Network pharmacology, transcriptomics and metabolomics were employed to identify the potential mechanism of GLBMY in TNBC treatment. Finally, the main regulating genes and proteins in the enriched pathways were validated by Quantitative real-time polymerase chain reaction (qPCR) and Western blotting analysis to confirm its mechanism. RESULTS: GLBMY can inhibit the growth of TNBC through apoptosis and necroptosis pathways and inhibit TNBC lung metastasis by inhibiting epithelial-mesenchymal transition (EMT). Network pharmacology has elucidated the most important active ingredients (tubeimoside I, emodin, cucurbitacin, and ursolic acid) and the most critical targets [interleukin-6 (IL6), signal transducer and activator of transcription 3 (STAT3), mitogen-activated protein kinase 3 (MAPK3)] of GLBMY in treating TNBC. RNA-seq revealed that GLBMY affected the nuclear factor kappa-light-chain-enhancer of activated B cells, rat sarcoma virus, and MAPK signalling pathways. Metabolomics revealed that the metabolites mainly affected by GLBMY were L-(+)-lactic acid, isocitric acid, benzoic acid and indoxyl sulfate. Subsequent experiments demonstrated that GLBMY can inhibit EMT in TNBC through the MAPK/ERK pathway and inhibit the proliferation and progression of TNBC through the IL6-STAT signalling pathway. CONCLUSIONS: We confirmed that GLBMY inhibits the development and metastasis of TNBC through the MAPK/Erk and IL6-STAT signalling pathways. GLBMY shows promise as a long-term supplementary or alternative therapy for TNBC, offering new insights for TNBC treatment.

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