Recent studies have revealed abnormal expression of miRNAs in various tumors. Although microRNA-338-3p (miR-338-3p) plays an important role in many types of tumors, its influence on liver cancer (LC) is unknown. In this study, we found that expression of miR-338-3p was decreased in LC cells and tissues. Colony formation and cell proliferation were suppressed by enhanced expression of miR-338-3p in LC cells. Moreover, miR-338-3p targeted sphingosine kinase 2 (SphK2). Silencing of SphK2 had an identical influence as overexpression of miR-338-3p in LC cells. Overexpression of SphK2 without the 3'-untranslated region remarkably enhanced the growth suppression triggered by miR-338-3p in LC cells. These findings indicate that miR-338-3p influences the development of LC by targeting SphK2, suggesting that miR-338-3p can be targeted as an innovative therapeutic strategy for LC.