Ganoderic Acid A Extract Induces Apoptosis and Upregulates Autophagy-Related Genes in NALM-6 Cell Line: A Potential Therapeutic Agent for Acute Lymphoblastic Leukemia

Abstract

Background: Acute lymphoblastic leukemia (ALL) is the most common leukemia in children and is associated with a high relapse rate despite current treatments. Ganoderic acid A (GAA) is a bioactive compound found in Ganoderma lucidum that has shown potential antileukemic properties. Objectives: This study aimed to investigate the effect of a GAA extract on autophagy gene expression and apoptosis induction in the NALM-6 cell line. Methods: A total of 20 × 103 NALM-6 cells were cultured in triplicate across various conditions: Culture condition supplemented with GAA, culture condition supplemented with L-asparaginase, culture condition supplemented with both GAA and L-asparaginase, and culture condition alone without L-asparaginase and GAA. The optimal concentration of GAA treatment was determined using an MTT assay. Flow cytometry was used to assess cell death induced by GAA treatment, using FITC-conjugated propidium iodide (PI) and annexin V staining. The expression levels of autophagy-related genes, including MAP1LC3B, BECN1, ATG5, ATG10, RB1CC1, and AMBRA1, were measured in the groups using real-time polymerase chain reaction. Results: The results of the MTT test indicated that the half maximum inhibitory concentration (IC50) of GAA against leukemic cells was 140 μg/mL after 48 hours of treatment. Moreover, flow cytometry analysis demonstrated a 40.5% increase in apoptosis and cell death at a concentration of 140 μg/mL of GAA after 48 hours. Furthermore, treatment with GAA resulted in upregulation of the expression of MAP1LC3B (P = 0.024), BECN1 (P = 0.035), ATG5 (P = 0.024), ATG10 (P = 0.024), RB1CC1 (P = 0.024), and AMBRA1 (P = 0.024) in NALM-6 cells compared to the control groups. Conclusions: These findings suggest that GAA has potential as a therapeutic agent for ALL. The GAA might have potential therapeutic properties against ALL.