Background <p>Lung cancer remains the leading cause of cancer-related mortality worldwide. Despite advances in therapeutic strategies, the 5-year survival rate for patients with non-small cell lung cancer (NSCLC) remains approximately 15%, highlighting the need for novel and more effective treatment approaches.</p> Objectives <p>The present study aimed to investigate the anticancer effects of a cannabidiol and bevacizumab combination in human NSCLC and to elucidate its underlying molecular mechanisms using the A549 cell line as an in vitro model.</p> Methods <p>A549 cells were cultured and treated with cannabidiol, bevacizumab, or their combination. Cell proliferation was assessed using the MTT assay, while apoptosis was quantified by flow cytometry. The expression of apoptosis-related proteins was evaluated by Western blotting, and the mRNA expression levels of metastasis-associated genes were analyzed using quantitative polymerase chain reaction (qPCR).</p> Results <p>Cannabidiol, bevacizumab, and their combination inhibited A549 cell proliferation in a dose-dependent manner. The apoptotic rate was 15.23% ± 0.42 following cannabidiol treatment and 21.97% ± 0.50 following bevacizumab treatment. Notably, combined treatment significantly increased apoptosis to 50.47% ± 0.67 (<i>p</i> &lt; 0.001). Western blotting demonstrated that co-treatment markedly downregulated the anti-apoptotic protein Bcl-2 and upregulated the pro-apoptotic protein caspase-9 (<i>p</i> &lt; 0.001). Furthermore, compared with either agent alone, combination therapy significantly reduced the expression of the metastasis-associated genes MMP-2 and MMP-9 (<i>p</i> &lt; 0.001).</p> Conclusion <p>The combination of cannabidiol and bevacizumab exerts a significantly enhanced anticancer effect on NSCLC cell growth inhibition compared with either agent alone, suggesting a promising therapeutic strategy for the management of NSCLC.</p> Graphical abstract <p></p>