Combination Treatment of Biochanin A and Atorvastatin Alters Mitochondrial Bioenergetics, Modulating Cell Metabolism and Inducing Cell Cycle Arrest in Pancreatic Cancer Cells

Abstract

Background/Aim: Pancreatic cancer is an aggressive type of cancer, with a dismally low survival rate of <5%. FDAapproved drugs like gemcitabine have shown little therapeutic success, prolonging survival by a mere six months. Isoflavones, such as biochanin A and daidzein, are known to exhibit anticancer activity, whereas statins reportedly have anti-proliferative effects. This study investigated the effects of combination treatment of biochanin A and atorvastatin on pancreatic cancer cells. Materials and Methods: Pancreatic cancer cells AsPC-1, PANC-1, and MIA PaCa-2 were procured from ATCC. The cell viability studies were carried out using MTT & cell count assays. Flow cytometry was used to study cell apoptosis whereas cell metabolism studies were carried out using the Seahorse Mito stress test and XF-PMP assay. The effects of treatment on cell signaling pathways & cell cycle associated proteins were investigated using western blot whereas invasiveness of cancer cells was evaluated using gelatin zymography. Results: The combination treatment decreased the survival and enhanced proapoptotic responses compared to single treatments in the pancreatic cancer cells. In PANC-1 cells, the combination treatment decreased invasiveness, reduced expression of activated STAT3 and expression of critical mediators of cell cycle progression. Furthermore, the combination treatment induced a differential inhibition of respiratory complexes in the pancreatic cancer cells. Conclusion: The combination treatment of biochanin A and atorvastatin exerts enhanced anti-cancer effects, inducing apoptosis, down-regulating cell cycle associated proteins and invasiveness in pancreatic cancer cells and merits further investigation for new, improved treatments for pancreatic cancer.Pancreatic cancer is the fourth leading cause of cancer-related deaths in the United States and has a very poor prognosis (1). In 2024, an estimated 66,440 new cases and 51,750 deaths are expected to be recorded due to this devastating disease (1). The poor prognosis associated with pancreatic cancer may be attributed to the limited effectiveness of current chemotherapies due to apoptosis resistance of cells in the primary tumor. The highly aggressive nature of the cancer cells and lack of biomarkers for early diagnosis of pancreatic cancer undoubtedly also contribute (2).Apoptosis resistance in pancreatic cancer has been attributed to several factors including impairment of TRAILmediated mechanisms promoting apoptosis and decreased TRAIL receptor expression (3-5). The low survival rate of pancreatic cancer can be mainly attributed to its highly aggressive nature. The tumor cells tend to metastasize to distant organs like the lungs and liver, which explains the primary tumor cells' resistance to anoikis (6-8). Some underlying mechanisms that promote resistance to anoikis and 2307