Wogonin Induces Apoptosis of Acute Myeloid Leukemia Cells through PI3K/Akt Pathway and Synergies with Venetoclax

Abstract

Background:Acute myeloid leukemia (AML) is a highly aggressive and heterogeneous hematological malignancy. Although 80% of patients with AML achieve complete remission after receiving standard chemotherapy, most relapse or die from treatment-related complications, and the overall prognosis remains less than 30%. Wogonin is a kind of natural flavonoid with anti-inflammatory, anti-tumor and other biological activities which could kill tumor cells by inducing cell cycle arrest. Venetoclax is a highly selective BCL-2 inhibitor that is currently approved for the treatment of chronic lymphocytic leukemia. Although venetoclax combined with demethylated drugs or low-dose cytarabine has become a first-line treatment option in elderly AML patients who were unable to tolerate high-intensity chemotherapy, resistance remained unavoidable, suggesting the need for more venetoclax-based combination regimenes. However, it was unclear whether Wogonin could promote the anti-AML effects of venetoclax. Objective: To explore whether Wogonin combined with venetoclax has synergistic effect in the treatment of acute myeloid leukemia and the potential synergistic mechanism. Methods: After treatment with Wogonin alone or combined with venetoclax, the viability of AML cell lines and primary AML cells was detected by cell proliferation assay, and the combined index of the two drugs was calculated by Compusyn software to evaluate the synergy effect. Apoptosis and cell cycle changes were detected by flow cytometry. The enrichment of differentially expressed genes and related signaling pathways in different drug treatment groups were analyzed by RNA sequencing. The expression levels of apoptosis-related proteins, BCL-2 family proteins, and the activation levels of PI3K/Akt signal related molecules were detected by western blot. Results: In AML cell lines and patient-derived primary tumor cells, Wogonin combined with venetoclax significantly inhibited cell viability, and the inhibitory effect on AML was stronger than that of the two drugs alone. Flow cytometry detected that Wogonin promoted apoptosis of AML cells in a dose-dependent manner and blocked the cells in the G0/G1 phase. Western blot verified that Wogonin promoted the cleavage of apoptotic proteins such as caspase3 and PARP, affected the expression of BCL-2 family proteins, such as down-regulating MCL-1 and BCL-xL, inactivating the PI3K/Akt/GSK3β signaling pathway, and further inducing the death of AML cells. Conclusion: Wogonin combined with venetoclax has a synergistic anti-AML effect, providing scientific basis for preclinical and clinical trials, and a potential combination strategy for the clinical treatment of AML.