Objective: Ovarian cancer remains one of the leading causes of mortality among gynecological malignancies. The OVCAR-3 cell line is commonly used as an in-vitro model in studies investigating ovarian cancer pathogenesis and therapeutic strategies. This study aimed to evaluate the cytotoxic effects of whey protein isolate using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and its antioxidant capacity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method. Materials and Methods: Buffalo milk was collected from the Guroymak district of Bitlis Province. Whey protein isolate was obtained and subsequently analyzed for antioxidant activity using the DPPH assay. Its anticancer effects were assessed on OVCAR-3 and HUVEC cell lines using the MTT assay. Experimental conditions included eight different concentrations (9.75, 19.50, 39, 78, 156, 312, and 624 µg/mL) together with a negative control group, and treatments were applied for 24 and 48 hours. Results: According to MTT analysis, the highest concentration (624 μg/mL) induced 18.54% cytotoxicity at 24 hours and 41.12% at 48 hours in OVCAR-3 cells (GraphPad analysis). A significant positive correlation was observed between buffalo milk and the resulting whey protein isolate (R² = 0.871, p 0.05). The DPPH assay demonstrated that whey protein isolated from buffalo milk exhibited notable antioxidant activity, with an IC₅₀ value of 93.64 μg/mL. Conclusion: In this study, whey protein isolate demonstrated measurable antioxidant activity and affected the viability of OVCAR-3 ovarian cancer cells in a dose- and time-dependent manner. The findings suggest that whey protein isolate may warrant further investigation to better understand its potential effects on cancer and normal cells in the context of antioxidant and anticancer research.
Sabancılar et al. (Sat,) studied this question.