Background: Tiny non-coding RNAs called microRNAs (miRNAs) are crucial for cell survival and differentiation. Numerous cancer varieties, including acute myeloid leukemia, have been linked to abnormal miRNA expression (AML). Objective: The objective of this study is to assess the prognostic importance of miRNA-181a-3p and its participation in the diagnosis of acute myeloid leukemia. Subjects and methods: Forty-five subjects were recruited from Benha University Hospital and Tanta Cancer Center. They were divided into 30 newly diagnosed adult patients with AML as patient's group and 15 apparently healthy individuals matching age and sex with patients as control group. At the beginning, blood samples (2 ml) were taken from the patients and control for complete blood picture and RT-PCR, and after 28 days of medication, more blood samples (1 ml) from the patient's group were taken in order to quantify the relative gene expression of miRNA-181a-3p. Results: The present study demonstrated a significant overexpression of miRNA -181a-3p gene as opposed to the control group in the AML group. Moreover, miRNA-181a-3p gene expression level decreased significantly after treatment when compared to its pretreatment level. Comparing post treatment level to control group, revealed that miRNA-181a-3p gene expression level decreased, but did not reach the control level. Non remission cases were significantly associated with higher baseline miRNA-181a-3p gene expression when compared to remitted cases. Conclusion: The current study revealed that miRNA-181a-3p expression level had a role in AML diagnosis and in prediction of prognosis.