Beyond Angiogenesis Inhibition: Bevacizumab-Induced Molecular Modulation and Immune Activation in A۵۴۹ Non-Small Cell Lung Cancer Cells

Background: The anti-tumor potential of bevacizumab, a monoclonal antibody that targets VEGF-A, can go beyond angiogenesis inhibition and can even lead to drug resistance. To seek further insights in resistance mechanisms to bevacizumab at the molecular level, we assessed gene expression and cytokine production in the human non-small cell lung cancer (NSCLC) cell line A۵۴۹. Methods: The A۵۴۹ cells were treated with different concentrations of bevacizumab for ۲۴ and ۴۸ hours. The MTT assay was performed to evaluate cell viability and morphology. Flow cytometry using Annexin V/PI staining was performed to quantify apoptotic and necrotic cell populations. The expression levels of p۵۳, Caspase-۳, and miR-۱۴۱ were quantified by real-time PCR, and fold changes were determined using the comparative Ct (ΔΔCt) method. Levels of IL-۶, IL-۸, and IP-۱۰ were measured by an ELISA test. Results: Bevacizumab significantly reduced cell viability in a dose- and time-dependent manner, with approximately ۴۰% viability at ۱۲.۵ µg/mL after ۴۸ hours. RT-PCR revealed increased expression of p۵۳ and miR-۱۴۱ with ۲.۱۲-fold and ۳.۳۰-fold, respectively. Caspase-۳ expression did not have a significant change. Flow cytometry result demonstrated that there was no significant change in the apoptotic activity. This suggests that the growth inhibition was not due to the classical apoptosis pathway. The cytokine analysis showed significant upregulation of IL-۶, IL-۸, and IP-۱۰ (p < ۰.۰۵), which can cause by activation of stress-related and immunemodulatory pathways. Conclusion: These findings show that bevacizumab has multi-level molecular effects in NSCLC. It can lead to a potential adaptive resistance and cellular senescence-like response through p۵۳ and miR-۱۴۱ activation. The rise in cytokine levels hints at possible immune involvement that could be used for therapeutic benefits. Our data strongly suggest consideration of molecular response profiles and cytokine activation for treatment, as well as designing novel combinatory therapies in the clinical setting.

کلیدواژه ها:

VEGF inhibition ، Non-small cell lung cancer ، Drug resistance ، A۵۴۹ cells ، Inflammatory response