〔Abstract〕 Objective To investigate the role of Tre2-Bub2-Cdc16 1 domain family member5(TBC1D5) in the development of hepatocellular carcinoma(HCC). Methods Western blot(WB), Immunohistochemistry(IHC) and quantitative real-time PCR(qPCR) were used to verify the difference inTBC1D5expression in clinical samples. The HCC cell lines MHCC97H and Hep3B were chosen to construct the knockdown model. The effects on cell proliferation were detected by cell proliferation assay, colony formation assay and EdU assay. Wound assay and Transwell assay were used to detect cell migration and invasion. Flow cytometry was used to detect the changes of cell cycle and H2O2-induced apoptosis of HCC cells. Finally, the effects of knockdown and overexpression ofTBC1D5on JAK/STAT pathway were detected by WB. Results The results of WB, IHC and qPCR showed that the expression ofTBC1D5in HCC tissues was higher at the protein level(P<0.000 1) and mRNA level(P<0.01) than that in corresponding adjacent tissues. Compared with the control group, the proliferation level of HCC cells withTBC1D5knockdown was decreased(P<0.05), the formation of plate colony number decreased(P<0.001), and the proportion of EdU-positive cells decreased(P<0.001). The results of scratch assay and Transwell assay showed that the migration(P<0.01) and invasion ability(P<0.01) of HCC cells afterTBC1D5knockdown were significantly lower than those in the control group. AfterTBC1D5knockdown, the cell cycle of HCC cells was slowed down(P<0.05) and the ability to resist apoptosis was reduced(P<0.01)than those in the control group. Compare with the control group, knockdown ofTBC1D5decreased the phosphorylation level of JAK and STAT proteins and inhibit the JAK/STAT pathway. Conclusion TBC1D5is highly expressed in HCC. After knocking downTBC1D5, the proliferation, migration and invasion ability, cell cycle rate and anti-apoptosis ability of HCC cells decreased and may affect HCC progression through the JAK/STAT pathway.