Herein, although we did not observe the associations of aberrant miR-335/ID4 expression with these gene mutations in AML, evidence shwed that miR-335/ID4 expression was associated Akt/pAkt expression in leukemic cell-lines and AML patients, and inhibition of PI3K/Akt signaling reversed and impaired the pro-proliferative and anti-apoptotic effects mediated by miR-335 overexpression in K562 and HL60 cell-lines. The gene discussed is AKT1; the disease is acute myeloid leukemia.