ABSTRACTHepatocellular carcinoma (HCC) is a major global health concern that accounts for more than 80% of all primary hepatic carcinomas. The long noncoding RNA FGD5 antisense RNA 1 (FGD5‐AS1) has been linked to HCC cell stemness and proliferation. However, the exact function of FGD5‐AS1 in HCC remains unclear. Cell viability and proliferation were examined using the CCK8 and colony formation assays, respectively. Cell stemness was examined using a sphere formation assay. To investigate the relation between Musashi 2 (MSI2) and FGD5‐AS1 (or protein kinase D1 [PKD1]), RNA immunoprecipitation and RNA pull‐down assays were used. Furthermore, a xenograft mouse model was established to evaluate the function of FGD5‐AS1 in vivo. FGD5‐AS1, MSI2, and PKD1 were upregulated in the HCC tissues. FGD5‐AS1 knockdown significantly inhibited the viability, proliferation, and stemness of HCC cells and decreased the expression of MSI2, PKD1, octamer‐binding transcription factor 4, SOX2, NANOG, and Prominin‐1 in HCC cells. Mechanistically, FGD5‐AS1 increased PKD1 mRNA stability by upregulating MSI2 expression. Both MSI2 and PKD1 ameliorated sh‐FGD5‐AS1's inhibition of HCC cell viability, proliferation, and stemness. Furthermore, FGD5‐AS1 silencing inhibited HCC tumor growth and stemness in vivo. FGD5‐AS1 promotes the stemness of HCC cells by activating the MSI2/PKD1 axis. Our study provides a new theoretical foundation for the development of novel HCC treatments.