Hesperidin increases the in vitro anticancer activity of Pentagamavunon-1 in triple‐negative breast cancer with limited effects on normal cells

Pentagamavunon-1 (PGV-1) is a potential anticancer candidate due to its specific targeting of cancer cells with minimal toxicity to normal cells. This study examined the potency of hesperidin (HSD) in improving the anticancer effects of PGV-1 for triple-negative breast cancer (TNBC). The 4T1 and NIH-3T3 cell lines were used as the model of TNBC and normal fibroblast, respectively. The cytotoxicity was quantified using MTT and clonogenic assays, whereas the cell cycle and apoptosis were measured by flow cytometry. Cell migration was assessed by scratch-wound healing assay. Senescence was measured using the SA-β-gal assay, and the secretion of matrix metalloproteinase (MMP)-2 and 9 was examined using gelatin zymography. Protein expression level was analyzed through capillary Western blot. The collected data were quantified using one-way ANOVA. PGV-1 significantly reduced 4T1 cell viability, with IC 50 values of 7 and 2 µM at 24 and 48 h, respectively. Its cytotoxic effect persisted up to 7 days after removal. HSD alone showed no cytotoxicity against 4T1 cells, but in combination with 1 µM of PGV-1, it synergistically enhanced cell growth inhibition and apoptosis induction. This synergism is likely due to ERK1/2 phosphorylation inhibition, leading to reduced levels of Bcl-2 and elevated levels of Bax. In NIH-3T3 cells, higher concentration PGV-1 (10 µM) induced early senescence-associated features, but 100 µM HSD prevented this and suppressed MMP-2 and 9 secretion. PGV-1 and HSD combination demonstrates potential as a co-treatment strategy for effective anticancer therapy while preserving normal cell integrity.