# Hesperidin Enhances Doxorubicin Efficacy by Modulating Apoptosis- and Migration-Associated Processes in Human Retinoblastoma Cells

## TL;DR

Hesperidin boosts the effectiveness of doxorubicin in treating retinoblastoma by increasing cell death and reducing cancer cell movement.

## Contribution

Hesperidin enhances doxorubicin's anti-tumor effects by modulating apoptosis and migration pathways in retinoblastoma cells.

## Key findings

- Hesperidin increases apoptosis and reduces cell migration when combined with doxorubicin.
- The combination downregulates MMP-2, MMP-9, and ACTA2 while upregulating TIMP-1 and TIMP-2.
- Apoptosis-related genes like Bax and Caspase-3 are upregulated, while Bcl-2 is downregulated.

## Abstract

Retinoblastoma (RB) is a rare pediatric eye cancer for which chemotherapy remains a cornerstone of treatment, although drug resistance and dose-related toxicity limit therapeutic success. Doxorubicin (DOX) is an effective anticancer agent, but its clinical utility can be improved by compounds that enhance its efficacy at lower doses. Hesperidin (Hes), a naturally occurring flavonoid, has attracted interest due to its biological activity in cancer-related cellular processes. In this study, we investigated whether Hes could enhance the cellular response to DOX in human RB cells under in vitro conditions. Our findings show that the combination treatment increases apoptosis, modulates extracellular matrix (ECM)-associated factors, and suppresses cell migration more effectively than either agent alone. These effects collectively contribute to an enhanced anti-tumor phenotype in RB cells. This work provides a preclinical framework supporting the potential of flavonoid-based chemosensitization strategies and highlights the need for further validation in advanced experimental models.

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.