Hong Hu, Chao Zhang, Zhiyong Liu
Objectives: To investigate the mechanism by which Hesperidin (Hsp) inhibits the proliferation and metastasis of the triple-negative breast cancer cell line MDA-MB-231. Methods: The effect of Hsp on cell viability 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 cell survival rate (p<0.01). The scratch test showed that Hsp inhibited wound healing in a concentration-dependent manner. The 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.