Comparison of energetic utilization of A549 human lung cancer cells at the level of allosteric glycolysis enzymes in 3D and 2D culture environments

Objective: İllustrate variations in glucose and glutamine utilization within A549 cancer cells cultivated using both 3D and 2D methods, focusing on the allosteric enzymes involved in glycolysis and the tricarboxylic acid (TCA) cycle. Materials and Methods: A549 cells were incubated using both 2D and 3D method in media containing either glucose, glutamine, or a combination of both for specific durations (24, 48, 72 hours). Following incubation, enzyme levels were measured in cell lysates using ELISA kits. MTT test was used for viability tests. For statistical analysis, the IBM SPSS Statistics 21 softeware was used. Results: In the glucose-glutamine containing medium, significant differences were observed in the concentrations of HK, PK, PDH, CS, IDH, and α-KGDC enzymes between 2D and 3D cultures at the 24th hour. In the glutamine medium, HK showed significant differences in both 2D and 3D cultures at all time intervals, while PFK exhibited higher concentrations in 3D cultures at the 24th and 72nd hours. Significant differences were observed in G6PDH and α-KGDC at the 48th and 72nd hours, and CS at the 24th and 72nd hours. In the glucose medium, significant differences were detected in HK, G6PDH, PFK, and IDH enzymes between 2D and 3D cultures at all time intervals. G6PDH, however, exhibited higher concentrations in the 2D method at all time intervals. Significant differences have been found in the concentrations of PK, PDH, CS, α-KGDC, and GDH enzymes between 2D and 3D culture methods. Conclusion: The disparities in cell viability and enzyme concentrations between 2D and 3D methods appear to be influenced by the incubation durations. These findings suggest that 2D and 3D culture methods have different effects on cellular metabolic activity, emphasizing the importance of detailed investigations in cell culture studies.