PURPOSE: Hesperidin, a glycoside flavonoid, is thought to act as an anti-cancer 
agent, since it has been found to exhibit both pro-apoptotic and 
anti-proliferative effects in several cancer cell types. The mechanisms 
underlying hesperidin-induced growth arrest and apoptosis are, however, not well 
understood. Here, we aimed to investigate the anti-proliferative and apoptotic 
effects of hesperidin on non-small cell lung cancer (NSCLC) cells and to 
investigate the mechanisms involved.
METHODS: The anti-proliferative and apoptotic effects of hesperidin on two 
NSCLC-derived cell lines, A549 and NCI-H358, were determined using a WST-1 
colorimetric assay, a LDH cytotoxicity assay, a Cell Death Detection assay, an 
AnnexinV-FITC assay, a caspase-3 assay and a JC-1 assay, respectively, all in a 
time- and dose-dependent manner. As a control, non-cancerous MRC-5 lung 
fibroblasts were included. Changes in whole genome gene expression profiles were 
assessed using an Illumina Human HT-12v4 beadchip microarray platform, and 
subsequent data analyses were performed using an Illumina Genome Studio and 
Ingenuity Pathway Analyser (IPA).
RESULTS: We found that after hesperidin treatment, A549 and NCI-H358 cells 
exhibited decreasing cell proliferation and increasing caspase-3 and other 
apoptosis-related activities, in conjunction with decreasing mitochondrial 
membrane potential activities, in a dose- and time-dependent manner. Through a 
GO analysis, by which changes in gene expression profiles were compared, we 
found that the FGF and NF-κB signal transduction pathways were most 
significantly affected in the hesperidin treated NCI-H358 and A549 NSCLC cells.
CONCLUSIONS: Our results indicate that hesperidin elicits an in vitro growth 
inhibitory effect on NSCLC cells by modulating immune response-related pathways 
that affect apoptosis. When confirmed in vivo, hesperidin may serve as a novel 
anti-proliferative agent for non-small cell lung cancer.