Evaluating the anticancer effects of carnosic acid against breast cancer: An In Vitro investigation.

Background: Carnosic acid (CA) has potential anti-cancer properties, but its effectiveness can be improved by combining it with Folic acid (FA). This research aimed to evaluate the impact of CA and CA-FA conjugate on breast cancer cell lines (MCF-7, MDA-MB-231, and MCA10).

Materials and methods: The viability of the cell lines was measured using the MTT assay, and the IC₅₀ was determined to compare the cytotoxicity of CA and CA-FA. The process of programmed cell death was investigated by utilizing Annexin V/PI staining, measuring caspase-3/7 activity, and real-time PCR for apoptotic gene expression. Reactive oxygen species (ROS) were also assessed to determine the extent of oxidative stress.

Results: CA significantly decreased the viability of MCF-7 and MDA-MB-231 cells depending on the dosage, with CA-FA exhibiting enhanced cytotoxicity, particularly in MDA-MB-231 cells. The evaluation of IC₅₀ confirmed that conjugation with FA reduced the IC₅₀ of CA. Apoptosis analysis demonstrated increased apoptosis rates in MCF-7 and MDA-MB-231 cells exposed to treatment with CA and CA-FA, while MCA10 cells showed minimal effects. Caspase-3/7 activity was notably higher in CA-FA-treated cells. Gene expression analysis revealed elevated pro-apoptotic gene activity and reduced anti-apoptotic gene activity, with CA-FA having a more pronounced effect. Cells subjected to CA-FA treatment exhibited a significant increase in ROS levels.

Conclusion: These findings suggest that CA conjugation with FA enhances its cytotoxic effects and promotes apoptosis through increased apoptosis and ROS production. The research emphasizes the promise of CA-FA as a focused treatment approach for aggressive forms of breast cancer, underscoring the need for additional exploration of its practical uses in clinical settings.