Cancer　stem-like　cells　(CSCs)　are　critical　for　the　initiation,　progression,　chemoresistance　and　postsurgical　recurrence　of　liver　cancer.　They　are　thought　to　be　novel　targets　for　the　treatment　of　liver　cancer,　however,　efficient　agents　that　target　liver　cancer　stem　cells　(CSCs)　have　not　been　identified.　MicroRNAs　(miRNAs)　are　small　non-coding　RNAs　that　target　the　3untranslated　region　(3UTR)　of　mRNAs.　Their　dysregulation　has　been　implicated　in　several　types　of　cancer　including　liver　cancer,　but　it　still　remains　unknown　if　they　play　a　role　in　targeting　liver　CSCs.　We　compared　the　miRNA　profiles　between　liver　cancer　samples　and　adjacent　non-tumor　tissues　using　The　Cancer　Genome　Atlas　(TCGA)　datasets.　Several　miRNAs　including　miR-486-5p　(miR-486)　were　found　to　be　significantly　downregulated　in　liver　cancer　tissues.　These　differentially　expressed　miRNAs　were　screened　between　CSC-enriched　tumor　spheres　and　adherent　cells.　miR-486　was　significantly　downregulated　in　tumor　spheres　and　liver　cancer　samples.　Ectopic　expression　of　miR-486　significantly　repressed　the　self-renewal　and　invasion　of　CSCs　in　vitro　and　tumorigenesis　in　vivo.　Notably,　we　found　that　sirtuin　1　(Sirt1)　served　as　a　direct　target　of　miR-486.　The　high　expression　of　Sirt1　was　involved　in　maintaining　the　self-renewal　and　tumorigenic　potential　of　liver　CSCs.　The　results　of　the　present　study　indicated　that　the　miR-486-Sirt1　axis　was　involved　in　suppressing　CSC　traits　and　tumor　progression.