Effect on MDA-MB231 human breast cancer cell line treated with bilberry (Vaccinium myrtillus) using Annexin V and AgNOR staining

Background/Aim: Cancer has become a prevalent disease, emerging as one of the major chronic health issues today. Currently, common treatments against cancer include chemotherapy, radiotherapy, surgery, and the use of chemically synthesized drugs. However, despite significant advancements in diagnostic methods and treatments, drug resistance and metastasis remain primary hurdles to successful cancer therapy. Consequently, attention has been shifted towards exploring alternative treatments and therapies against cancer. This study sought to examine the time and dose-dependent effects of blueberry (Vaccinium myrtillus L) on MDA-MB-231 cell lines. Methods: The study used the MDA-MB231 breast cancer cell line. We established three groups: control, 40 µl/ml bilberry, and 80 µl/ml bilberry, which were incubated at 37°C and 5% CO2 for 24 and 48 h, respectively. After incubation, we examined the viability, apoptosis, and cell cycle of MDA-MB-231 cells with the Muse Cell Analyzer and assessed the status of nucleolar organizer region (NOR) proteins via silver nitrate (AgNOR) staining. Results: Bilberry extracts were found to enhance apoptosis and exhibit a cytotoxic effect, thereby reducing cell proliferation in MDA-MB-231 cells after 24 and 48 h of culture. There was notably increased apoptosis at concentrations of 40 µl and 80 µl. Moreover, after 48 h of incubation, a significant difference emerged between the control and 40 µg/ml bilberry samples, notably in the average AgNOR count and the total AgNOR area/total nuclear area ratio. Conclusion: Our study suggests that blueberries may be a potential therapeutic candidate for cancer treatment, thereby potentially enriching cancer research.