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Objective To investigate the impact of the RNA-binding protein with multiple splicing(RBPMS), which has multiple splicing functions, on the function of acute myeloid leukemia cells line THP-1 carrying the MLL-AF9 fusion gene. Methods The expression of RBPMS in the hematopoietic system was analyzed using the GEO database; RBPMS over-expression in THP-1 cells was achieved through lentiviral infection. The efficiency was detected by RT-qPCR and Western blot; The effect of RBPMS over-expression on the THP-1 cell transcriptome was evaluated by high-throughput sequencing. The impact of RBPMS over-expression on the proliferation and migration of leukemia cells was examined using CCK-8 assay and flow cytometry, respectively. Results RBPMS expressed in acute myeloid leukemia stem cells carrying the MLL-AF9 fusion gene was found to be inhibited. Over-expression of RBPMS significantly reduced the proliferation and migration of THP-1 cells. Conclusions RBPMS inhibits cell proliferation and migration of acute myeloid leukemia cells line carrying the MLL-AF9 fusion gene.