Taraxerol targets the HIF1A-NEK6 axis to block TDE-mediated M2 macrophage polarization and inhibits NSCLC progression
Qiang Yang1, Hongli Li2, Baojun Guo1
1Department of TCM, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Zhengzhou, Henan 450003, China.
Background:
Non-small cell lung cancer (NSCLC) progression is tightly linked to tumor microenvironment (TME) remodeling. Taraxerol, a natural compound, has shown potential antitumor activity, but its molecular mechanisms in regulating NSCLC, particularly in TME-related progression, remain poorly characterized.
Methods:
Single-cell RNA sequencing (scRNA-seq) was conducted to characterize cellular heterogeneity. Network pharmacology integrated differential gene analysis and taraxerol target prediction to identify key regulatory nodes. Molecular interactions were verified via molecular docking and molecular dynamics (MD) simulations. CCK-8, EdU, flow cytometry, and Transwell were employed to assess cell proliferation, apoptosis, migration, and invasion. Xenograft models were established to elucidate taraxerol's function in vivo. Subsequently, tumor-derived exosomes (TDEs) were isolated and characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot. Macrophage characteristics were validated via qRT-PCR. The interaction between hypoxia-inducible factor 1 alpha subunit (HIF1A) and NIMA-related kinase 6 (NEK6) was investigated via chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays.
Results:
scRNA-seq revealed NSCLC-specific cellular remodeling, with distinct heterogeneity in macrophage subsets and altered proportions of tumor epithelial cells. HIF1A was a core target of taraxerol, confirmed by molecular docking (binding affinity -8.5 kcal/mol) and MD simulations showing stable interactions. Taraxerol inhibited HIF1A expression in A549/H1299 cells, suppressing proliferation, migration, invasion, and in vivo NSCLC tumorigenesis while promoting apoptosis. Taraxerol-treated TDEs reduced M2 polarization and migration of macrophages, while HIF1A overexpression restored these effects. Furthermore, HIF1A transcriptionally activated NEK6 by directly binding to its promoter, whereas taraxerol blocked the HIF1A-NEK6 axis-mediated promotion of macrophage M2 polarization and migration.
Conclusion:
Taraxerol exerted antitumor effects in NSCLC by targeting HIF1A, thereby regulating TDE-mediated macrophage M2 polarization and tumor progression via the HIF1A-NEK6 axis.
Insights
Taraxerol, a natural compound, inhibits non-small cell lung cancer (NSCLC) progression by targeting HIF1A. This mechanism regulates tumor microenvironment remodeling, specifically macrophage polarization, via the HIF1A-NEK6 axis.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Non-small cell lung cancer (NSCLC) progression is influenced by tumor microenvironment (TME) remodeling.
- Taraxerol shows potential antitumor activity, but its mechanisms in NSCLC TME regulation are unclear.
Purpose of the Study:
- Investigate Taraxerol's molecular mechanisms in NSCLC, focusing on TME interactions.
- Elucidate Taraxerol's role in regulating macrophage polarization and NSCLC progression.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) for cellular heterogeneity analysis.
- Network pharmacology, molecular docking, and simulations to identify Taraxerol's targets.
- In vitro assays (proliferation, apoptosis, migration, invasion) and in vivo xenograft models.
- Analysis of tumor-derived exosomes (TDEs) and macrophage polarization.
- Investigation of the hypoxia-inducible factor 1 alpha subunit (HIF1A)-NIMA-related kinase 6 (NEK6) interaction.
Main Results:
- scRNA-seq identified NSCLC-specific remodeling, including macrophage heterogeneity.
- HIF1A was identified as a core target of Taraxerol, with stable binding confirmed by simulations.
- Taraxerol suppressed NSCLC cell proliferation, migration, invasion, and tumorigenesis, while promoting apoptosis.
- Taraxerol-treated TDEs reduced M2 macrophage polarization and migration; HIF1A overexpression reversed this.
- Taraxerol blocked the HIF1A-NEK6 axis, inhibiting M2 macrophage polarization and migration.
Conclusions:
- Taraxerol exhibits antitumor effects in NSCLC by targeting HIF1A.
- Taraxerol regulates TDE-mediated macrophage M2 polarization and NSCLC progression.
- The HIF1A-NEK6 axis is a key pathway targeted by Taraxerol in NSCLC.
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