Farnesol suppresses hypoxia associated EMT signaling and impairs 3D spheroid integrity in A549 lung adenocarcinoma cells.

Lung cancer remains the leading cause of high cancer incidence and mortality worldwide. Among lung cancers, non-small cell lung cancer (NSCLC) is the predominant subtype accounting for majority of cancer deaths. The invasive and metastatic behavior of NSCLC largely contributes to its aggressive nature and therapeutic resistance. Farnesol (FAR), a naturally occurring sesquiterpene, has shown anticancer activity in several malignancies however, its mechanistic role in NSCLC invasion and metastasis remains not fully understood. In the present study, we investigated the effect of farnesol on A549 lung adenocarcinoma cells through in vitro approaches involving protein-protein/gene interactions, molecular analysis and 3D screening. Scratch assay and transwell invasion assays were performed to evaluate the effect of farnesol on A549 cell migration and invasion. Immunofluorescence staining analysis were employed to assess the anti-angiogenic potential of farnesol followed by western blotting and real-time PCR to validate the efficacy of farnesol in regulating invasion and metastasis associated markers. Farnesol significantly reduced the viability of A549 cells in a dose-dependent manner, with an IC₅₀ of 21.5 µg/mL inducing cell shrinkage and loss of cell density. Scratch and matrix-assisted transwell invasion assays demonstrated marked inhibition of migration and invasion with increased nonmigratory spaces in farnesol-treated group compared to control. Molecular analysis confirmed upregulation of E-cadherin, downregulation of VEGF, PCAF, COX2, and suppression of mesenchymal and matrix remodeling markers. Farnesol reduced spheroid diameter, dissociation of spheroid integrity and induced of apoptosis at concentrations above 125 µg/mL. Collectively, our findings suggest that farnesol potentially inhibits A549 cell migration, invasion and metastasis, highlighting farnesol as an effective therapeutic candidate for the treatment of lung adenocarcinoma.