2‐Pyrazoline‐5‐One Derivative Suppresses Cell Proliferation and Induces Apoptosis With Upregulation of Bax in Breast Cancer Cells

ABSTRACT

The most common cancer in women worldwide is still breast cancer. Among its subtypes, luminal A and triple‐negative breast cancers present significant therapeutic challenges due to intrinsic drug resistance and absence of effective targeted treatments. The pharmacological potential of heterocyclic compounds has garnered more attention in recent years, and structures like pyrazolines, hydrazones, and triazoles are important in the development of new drugs. Compounds incorporating these scaffolds have been shown in numerous studies to have strong anticancer action. In this regard, pyrazoline derivatives have become a viable field of study for focused treatment approaches. This study focused on evaluating the cytotoxic potential of the 2‐pyrazoline‐5‐one derivative compound against luminal‐A (MCF‐7) and triple‐negative (MDA‐MB‐231) breast cancer cell lines. The CCK‐8 test was used to assess the impact on cell viability. Additionally, Annexin V/PI staining was used to identify the apoptotic effects, and fluorescence microscopy was used for analysis. The wound healing experiment was used to measure the capacity of cells to migrate, and migration process was monitored following scratch formation. The colony formation assay was used to assess the potential for cell proliferation, and clonal growth capacities were compared. Bax protein levels in compound‐treated cancer cells were quantified by ELISA to investigate the apoptotic mechanism. IC50 values in MCF‐7 and MDA‐MB‐231 cell lines were used to evaluate the cytotoxic effects of the 2‐pyrazoline‐5‐one derivative. The compound exhibited strong cytotoxic activity and reduced cell viability in both cell lines in a concentration‐dependent manner, with IC ₅₀ values of 13 µM in MCF‐7 cells and 16 µM in MDA‐MB‐231 cell. Furthermore, the compound effectively induced apoptosis and significantly suppressed colony formation and cell migration in both cell lines. Additionally, it showed a concentration‐dependent rise in Bax protein levels, suggesting a pro‐apoptotic action. These results confirm that the 2‐pyrazoline‐5‐one derivative has strong antiproliferative and pro‐apoptotic activity through Bax activation, providing a possible pathway for the creation of targeted treatments for breast cancer.