Exploring the mechanism of feiyanning formula and its extract apigenin against EGFR-TKIs resistance in non-small cell lung cancer based on UPLC-HRMS and experimental validation

Abstract

Ethnopharmacological relevance: Drug resistance in lung cancer poses a significant challenge in clinical treatment. Feiyanning formula (FYN) is a traditional Chinese medicine formula developed through years of clinical practice. However, the specific mechanisms by which FYN and its extract inhibit drug resistance in lung cancer remain unclear.

Aim of the study: This study aims to elucidate the therapeutic potential and underlying molecular mechanisms of FYN and its extract in the treatment of lung cancer.

Methods: To explore how FYN interacts with potential targets in treating non-small cell lung cancer (NSCLC), a comprehensive strategy was employed, including network pharmacology, ultrahigh-performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-HRMS), cellular assays, animal models, and molecular docking. Protein-protein interaction (PPI) network construction and functional analysis were used to investigate how FYN and its extract, apigenin, may overcome resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs). The molecular mechanisms were further validated using an NSCLC resistance animal model.

Results: The study confirmed that FYN and its extract inhibit the proliferation, migration, and invasion of H1975OR cells. Using network pharmacology and UPLC-HRMS, shared components between the FYN formula and drug-containing serum were identified, and a potential target network related to NSCLC resistance was constructed. Gene enrichment analysis indicated that FYN targets the IGF1R-PI3K-Akt signaling pathway in combating NSCLC resistance. Molecular docking revealed interactions between FYN's extract and key pathway-related genes. Cell phenotype assays showed that FYN not only suppresses proliferation, migration, and invasion of resistant NSCLC cells but also induces apoptosis and promotes cell cycle arrest. Additionally, animal experiments demonstrated that FYN significantly inhibits the growth of H1975OR xenografts in nude mice. Western blot analysis suggested that FYN may suppress the IGF1R-PI3K-Akt pathway in vivo.

Conclusions: This study explores the pharmacological effects and underlying mechanisms of FYN and its extract, apigenin, in the treatment of drug-resistant NSCLC. The findings support the therapeutic efficacy of FYN as a potential clinical treatment for lung cancer and provide robust scientific evidence for further investigation into the mechanisms of osimertinib resistance and the action of FYN.

Keywords: Apigenin; EGFR-TKIs resistance; Feiyanning formula; IGF1R-PI3K-Akt pathway; Non-small cell lung cancer.