Objective To investigate the expression of circAMOTL1 in liver cancer cell lines and its effect on the proliferation, migration and malignant transformation of liver precursor cell line WB-F344. Methods The expression of circAMOTL1 in five liver cancer cell lines, normal liver cell line L-02 and hepatic progenitor cell line WB-F344 was detected by real-time fluorescence quantitative PCR. The WB-F344 cell line with overexpression or knockdown of circAMOTL1 was constructed separately. CCK-8 assay was used to detect the proliferation of hepatic progenitor cells after circAMOTL1 overexpression or knockdown. Cell scratch test was used to verify the changes in cell migration. Western blot was used to verify the effect of overexpression or knockdown of circAMOTL1 on the expression of liver cancer stem cell marker CD133. Results The expression level of circAMOTL1 in the five HCC cell lines was higher than that in L-02(P＜0.05) The proliferation ability of hepatic progenitor cells was significantly increased after circAMOTL1 overexpression (P＜0.01), and the cell proliferation ability was significantly decreased after knocking out circAMOTL1 (P＜0.001). Overexpression of circAMOTL1 significantly promoted cell migration (P＜0.01), knockout (P＜0.01), the migration ability of the cells was significantly decreased (P＜0.01). CD133 expression was detected after circAMOTL1 overexpression. Conclusion circAMOTL1 was highly expressed in five HCC cell lines. Overexpression of circAMOTL1 can promote the proliferation and migration ability of hepatic progenitor cells and induce their malignant transformation into liver cancer stem cells