Azenosertib: wee1 inhibitor's anti-cancer effect determination on triple negative breast cancer

INTRODUCTION AND OBJECTIVE: Cell cycle checkpoints play a fundamental role in ensuring the necessary repair and genomic integrity by stopping the cycle in the presence of DNA damage. Wee1 kinase, which has significant activity at cell cycle checkpoints, is overexpressed in many types of cancer. In this context, treatments startegies targeting Wee1 inhibition have become prominent recently. This study aims to determine, for the first time, the potential therapeutic efficacy of Azenosertib as a Wee1 inhibitor in triple negative breast cancer (TNBC) at the molecular level. MATERIALS AND METHODS: The cytotoxic effect of Azenosertib on MDA-MB-231 TNBC and MCF-10A control cells was determined by CCK-8 analysis. Its apoptotic effect and activity in the cell cycle were determined by Annexin V analysis, acridine orange/propidium iodide staining and cell cycle analysis. Mitochondri staining and ROS analysis were conducted to determine the antioxidant effect of Azenosertib on MDA-MB-231 cells. Finally, the effect of Wee1 inhibition on the cell cycle was elucidated by western blot analysis. RESULTS: Azenosertib significantly reduced MDA-MB-231 cells viability time and concentrations-dependent (p<0.01). Although it caused apoptotic death in cells, its effect on the cell cycle was determined by increasing in Wee1 protein level depending on the incubation time. Additionally, Azenosertib caused mitochondrial damage and an increase in the amount of intracellular ROS in TNBC cells. CONCLUSION: The potential anticancer activity of Azenosertib in TNBC cells was determined, for the first time, at the molecular level. The findings showed that Azenosertib caused apoptosis and oxidative damage by inhibiting Wee1 activity in TNBC cells. New studies can be carried out to investigate the therapeutic effectiveness of Azenosertib, which was shown to be effective on TNBC cells for the first time, in other types of cancer and different types of death as well as apoptosis.