Abstract
This study evaluated the therapeutic potential and underlying molecular mechanisms of the citrus flavonoid Hes in combination with the chemotherapeutic agent DOX in human RB cells. Cells were treated with Hes alone or in combination with DOX for 24 and 48 h. Hes significantly inhibited cell proliferation and migration and promoted apoptotic cell death, while enhancing the cytotoxic response to DOX under in vitro conditions. Molecular analyses demonstrated that combination treatment markedly modulated ECM-associated markers, including the downregulation of matrix metalloproteinases MMP-2 and MMP-9 and ACTA2 (α-smooth muscle actin, α-SMA), along with the upregulation of tissue inhibitors of metalloproteinases TIMP-1 (tissue inhibitor of metalloproteinases) and TIMP-2. In parallel, the expression of apoptosis-related genes was altered, as evidenced by the upregulation of the B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax) and Caspase-3 and the downregulation of Bcl-2. Overall, these findings indicate that Hes enhances DOX efficacy by simultaneously engaging apoptotic and migration-associated molecular processes, supporting its potential role as a preclinical chemosensitizing agent that warrants further investigation in advanced experimental models.