In our in vivo study, tumor growth was inhibited after intratumoral injection of cholesterol-conjugated miR-541-5p inhibitor but increased by intratumoral injection of cholesterol-conjugated circMTO1 siRNA, indicating that the in vivo intervention of circMTO1/miR-541-5p/ZIC1 axis could be a potential target in HCC targeted therapy. This evidence concerns the gene ZIC1 and hepatocellular carcinoma.