Isoginkgetin inhibits non-small cell lung cancer by inducing oxidative stress and regulating M1 macrophage polarization

Background: Non-small cell lung Cancer (NSCLC), which constitutes approximately 85% of all lung Cancer cases, is a leading cause of Cancer mortality worldwide. Isoginkgetin (ISO), a biflavonoid isolated from Ginkgo biloba leaves, has demonstrated Anticancer effects against various malignancies. However, its anti-NSCLC effects are unclear.

Objective: To explore the therapeutic potential of ISO and its mechanism of suppressing NSCLC proliferation. To provide experimental evidence for the translational potential of ISO as a candidate clinical drug.

Methods: Cell cytotoxicity and proliferation were assessed by cell viability, propidium iodide staining, colony formation, and cell cycle detection assays. Cell migration was evaluated by wound healing and transwell assays. Reactive Oxygen Species and the mitochondrial membrane potential were detected by fluorescent staining. Malondialdehyde and glutathione content were quantified by commercial assay kits. Transcriptome analysis for exploring the mechanism of ISO inhibition in NSCLC. The relationships between ISO and the tribbles pseudokinase 3 (TRIB3)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway were investigated through Western blot, siRNA-mediated TRIB3 knockdown, and overexpression of TRIB3. Cellular thermal shift assay and surface plasmon resonance were explored to investigate the binding and direct interaction between ISO and TRIB3. A non-contact co-culture model of NSCLC cells and macrophages was used to assess macrophage polarization. The anti-NSCLC effect of ISO in vivo was evaluated in a BALB/c nude mouse subcutaneous tumor model.

Results: ISO suppressed the cell viability, proliferation, and migration of NSCLC cells. ISO induces oxidative stress in NSCLC cells, characterized by an increase in Reactive Oxygen Species and malondialdehyde and a decrease in mitochondrial membrane potential and glutathione. ISO has a strong binding affinity and direct interaction for the TRIB3 protein. Furthermore, ISO inhibited the antioxidant Nrf2 pathway by suppressing TRIB3 expression. Mechanistically, TRIB3 knockdown or overexpression significantly regulated the levels of oxidative stress and the Nrf2 pathway. ISO also promoted M1 macrophage polarization. Furthermore, ISO suppressed tumor growth in vivo.

Conclusion: ISO inhibits NSCLC through direct targeting and downregulation of TRIB3, thereby inducing oxidative stress and suppressing the antioxidant Nrf2 pathway. Furthermore, ISO regulates the tumor microenvironment by promoting M1 macrophage polarization. These findings may contribute to improved prognosis for NSCLC patients and provide insights into the mechanism of ISO as an antitumor drug, thereby supporting its potential for clinical translation.

Isoginkgetin; Non-small cell lung cancer; Nrf2; Oxidative stress; TRIB3.