Exploration of the mechanism of hesperidin inhibition on the proliferation and metastasis of human breast cancer cells.

OBJECTIVE: To investigate the mechanism of Hesperidin (Hsp) in inhibiting the proliferation and metastasis of breast cancer cell line MDA-MB-231. METHODS: The effect of Hsp on cell survival rate was analyzed using the MTT assay. The anti-migration and anti-invasion abilities of Hesperidin were evaluated using the scratch test and Transwell migration assay. The activity of matrix metalloproteinases (MMP)-2/MMP-9 was analyzed using gelatin zymography. The expression of epithelial-mesenchymal transition (EMT)-related proteins (Vimentin, Snail, ZO-1) was detected by Western blot. RESULTS: Low concentrations of Hsp (2-10 μmol/L) showed no significant cytotoxicity; 20-40 μmol/L significantly reduced survival rate (P<0.01). Scratch test results showed that Hsp inhibited wound healing in a concentration-dependent manner. Transwell migration assay showed that the number of migrating cells decreased with increasing Hsp concentration. Gelatin zymography results indicated that MMP-2/MMP-9 activity decreased with increasing Hsp concentration; Western Blotting results showed that Hsp downregulated the metastasis-related proteins Vimentin and Snail and upregulated the adhesion protein ZO-1. CONCLUSION: Our findings suggest that Hsp inhibits tumor cell invasion and metastasis by potentially reducing MMP-2/MMP-9 hydrolase activity, blocking extracellular matrix degradation, and reversing the EMT process (downregulating Vimentin/Snail and upregulating ZO-1). Hsp may represent a promising candidate for adjuvant therapy in breast cancer.