Cancer stem cells (CSCs) are related to tumorigenesis, recurrence, metastasis, and drug resistance in hepatocellular carcinoma (HCC).1 Let-7i is a noncoding small RNA, belonging to the famous miRNA let-7 family. Recently, let-7i has been revealed as a tumor suppressor on CSCs in ovarian cancer.2 However, let-7i is overexpressed in side population cells of rat HCC.3 It is unknown about the role of let-7i on human CSCs in HCC. Wnt pathway has critical roles in tumorigenesis, including differentiation, proliferation, and drug resistance of liver cancer. Let-7i inhibits osteogenesis by targeting Wnt pathway.4 Moreover, let-7i-3p suppresses cell growth and β-catenin in hepatoblastoma.5 Nevertheless, the effect of let-7i-5p on Wnt pathway needs to be uncovered in HCC. In this study, we aimed to investigate the effect and mechanism of let-7i-5p on stemness maintenance of CSCs in HCC.