Antioxidant activity and cytotoxic effects of Benincasa hispida juice on HepG2 cells

Benincasa hispida Cogniaux is widely utilized as a dietary staple and a medicinal remedy in China owing to its diverse biological properties. However, its potential antioxidant and anticancer properties has rarely been systematically reported. Therefore, this study aimed to investigated the in vitro antioxidant and anticancer effects of Benincasa hispida juice (BHJ). The total phenolic and flavonoid contents of BHJ were determined to be 37.14 ± 4.26 mg GAE/L and 12.43 ± 2.36 mg RUT/L juice, respectively. BHJ at the original concentration (1×) exhibited potent scavenging activity against DPPH radical, superoxide anions(O₂^-), hydroxyl radicals(·OH) and nitrite, with respective scavenging rates of 43.15 ± 2.18%, 24.35 ± 2.37%, 19.57 ± 1.49% and 38.64 ± 3.42%. Treatment of HepG2 cells with BHJ at concentration of 10, 20, and 40 mL/L significantly induced cellular apoptosis, which was accompanied by upregulation of activated caspase 9, PARP, Bax, Bad, and Apaf-1, as well as downregulation of Bcl-2 and pro-caspase 3. Furthermore, caspase-specific inhibitors (Ac-DEVD-CHO and Z-LEHD-FMK) effectively abrogated BHJ- induced reduction in HepG2 cells viability. Additionally, BHJ at 5 and 10 mL/L significantly suppressed the migration and invasion of HepG2 cells. This anti-metastatic effect was associated with the downregulation of matrix metalloproteinase(MMP)-2 and MMP-9, along with the upregulation of TIMP-1. Collectively, these findings demonstrate the potent antioxidant and anticancer capacities of BHJ, suggesting that Benincasa hispida may serve as a promising source of therapeutic agents for combating oxidative stress and hepatocellular carcinoma.