Study on the Improvement of Immune Microenvironment in Lung Cancer by the Combination of Three-Step Analgesics and Glabrene Based on Network Pharmacology and Molecular Docking Analysis

Abstract

Background: The treatment of lung cancer continues to encounter substantial challenges, necessitating ongoing exploration of effective drugs and targets. This study aims to investigate the regulatory mechanism behind the combined application of the three-step analgesic morphine and the active ingredient Glabrene found in the Tianchan capsule, within the context of non-small cell lung cancer (NSCLC). Methods: A549 cells were treated with morphine (10 μM) and morphine combined with Glabrene (3.5 μM) for 48 h. Cell proliferation was detected using 5-ethynyl-2′-deoxyuridine (EdU) staining. Alterations in cell invasion capacity were evaluated using the Transwell® assay. The expression levels of fibroblast growth factor receptor 3 (FGFR3), Vimentin, and E-cadherin were measured using quantitative polymerase chain reaction (qPCR). Subsequently, A549 cells were implanted into the left axillary region of nude mice. The mice were treated with morphine (1.5 mg/kg) or Glabrene (0.5 mg/kg) or morphine combined with Glabrene (0.5 mg/kg). Tumor mass changes were evaluated after 4 weeks. Finally, network pharmacology and molecular docking analysis were used to explore the relationship between Glabrene and FGFR3. Results: In this study, the results indicate that the combination of morphine and Glabrene effectively inhibits tumor growth in the NSCLC mouse model and reduces cancer cell proliferation and invasion (p < 0.05). Network pharmacology and molecular docking analysis identified Glabrene, the active ingredient found in the Tianchan capsule, as well as its target gene, FGFR3. Bioinformatics analysis revealed a compelling association between FGFR3 and numerous immune pathway marker genes. Moreover, FGFR3 exhibited a significant correlation with the immune infiltration scores (p < 0.05). qPCR testing showed that the expression levels of FGFR3 and Vimentin in the morphine + Glabrene group and the Glabrene group were significantly lower than those in the control group and the morphine group (p < 0.05). Conclusions: The combination of morphine and Glabrene or Glabrene alone can significantly inhibit tumor growth and reduce cancer cell proliferation and invasion in the NSCLC mouse model. Glabrene exhibits a strong affinity for FGFR3, which is significantly associated with immune pathways and immune infiltration.