Burkitt lymphoma (BL) is a highly aggressive B‐cell lymphoma that includes two forms of BL differing in Epstein–Barr virus (EBV) infection status, EBV‐positive and EBV‐negative. Although many efforts, such as high‐intensity, short‐duration combination chemotherapy, have been devoted to improving therapy for this rapidly proliferating neoplasm, there are still significant treatment‐associated toxicities. Therefore, there remains a need for novel effective therapeutic strategies. MicroRNAs play a role in “fine tuning” the physiological and pathological differentiation process, by which cells can rapidly regulate dynamic events such as cell‐lineage decisions and morphogenesis. This unique miRNA feature shifts the traditional one drug target paradigm to a novel one drug multiple targets paradigm. Here, we found that BL cell lines showed an extremely low expression of microRNA‐150 (miR‐150), and then restored miR‐150 expression at physiologic levels in BL cell lines Daudi, Raji, BJAB, and Ramos. The results showed that re‐expression of miR‐150 reduced proliferation of Daudi and Raji cells. Furthermore, Daudi and Raji, both of which are of EBV‐positive germinal center B‐cell origin, transduced with miR‐150 can be rescued to differentiate toward B‐cell terminal stage. However, no significant changes were observed in BJAB or Ramos cells, which are of EBV‐negative germinal center B‐cell origin. Of note, re‐expression of miR‐150 also resulted in decreasing c‐Myb protein levels. Additionally, c‐Myb knockdown in Daudi and Raji cell lines recapitulated the partial characteristics similar to that caused by re‐expression of miR‐150. Taken together, our findings show that miR‐150 can induce EBV‐positive BL differentiation by targeting c‐Myb.