Abstract

Effects of L-asparaginase on The Expression of EBF1 Gene, miR-181b, and miR-17 and Its Association with Apoptosis in the Nalm-6 Cell Line

Background: L-asparaginase is one of the most important and common drugs in the treatment of children and adults with acute lymphoblastic leukemia. Transcription factors play a key role in the prognosis, treatment, and recurrence of cancer. The EBF1 transcription factor is essential for B cell commitment and differentiation. Moreover, miRNAs are involved in many cellular processes such as cancer. Aim of the work: This study aimed to evaluate the effect of L-asparaginase on the EBF1 gene, miR-181b, and miR-17 expression and to examine its relationship with apoptosis in the Nalm-6 cell line. Material and Methods: In this experimental study, the Nalm-6 cell line was cultured in optimal condition, and treated with L-asparaginase. An ideal concentration of L-asparaginase for the Nalm-6 cell was determined by using the MTT assay. Before and after treatment with L-asparaginase, Apoptosis was evaluated by using flow cytometry, also the expression of EBF1 gene, miR-181b, and miR-17 was performed by real-time PCR assay. Results: The results showed significant percentages of apoptotic cells at 48 (34.35%) and 72 hours (67.30%) after treatment with L-asparaginase. There was no significant difference in the EBF1 gene expression before and after treatment (p<0.07). The expression of miR-181b (p<0.0001) and miR-17 (p<0.0001) was significantly decreased in the treated Nalm-6 cells as compared to the untreated Nalm-6 cells. Conclusion: After treatment Nalm-6 by L-asparaginase, there was a remarkable increase in apoptosis which was time-dependent. The expression of miR-181b and miR-17 was also notably reduced but there was no significant change in EBF1gene expression.

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