Background: Acute lymphoblastic leukemia (ALL) is a malignant disorder in both humans and animals. L-asparaginase (LASNase) has limitations as a chemotherapy agent due to adverse effects and low serum stability. In a previous study, L-ASNase was chemically modified with carboxymethyl dextran to enhance its properties. Aims: This study aimed to validate the potential of these modifications using the NALM-6 cell line. Methods: NALM-6 cells were cultured and treated with various concentrations, including 0 IU/ml as negative control, 0.5, 1, 1.5, and 2 IU/ml of modified L-ASNase and L-ASNase. The optimal concentration was determined at specific intervals, and viability and metabolic activity were assessed through Trypan blue and MTT tests. Flow cytometry, using Annexin V/PI staining, was employed to evaluate apoptosis. Real-time RT-PCR techniques were used to determine changes in the expression of the ATG2B and LC3-II genes (important genes in autophagy), with data analysis conducted using PRISM software. Results: The modified L-ASNase reduced the viability of NALM-6 cells and induced higher levels of apoptosis (P=0.001). Interestingly, the modified enzyme had a lesser impact on autophagy, which is important for avoiding treatment resistance. Conclusion: The modified L-ASNase showed enhanced effectiveness in reducing the viability of NALM-6 cells and induced higher levels of apoptosis. Interestingly, the modified enzyme had a lesser effect on autophagy, which is important as excessive autophagy can lead to treatment resistance. These findings suggest that the modified L-ASNase may have the potential to be a more effective chemotherapeutic agent for ALL treatments.

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