Abstract: To determine the role and mechanism of DEP domain containing 1（DEPDC1） in regulating hepatocellular carcinoma（HCC） cells glycolysis, differentially expressed genes（DEGs） in Huh-7 cells which DEPDC1 has been knockdowned were screened. The DEGs were analyzed by gene enrichment analysis（GSEA） to find significantly enriched gene sets and signaling pathways. Western blot was used to detecte DEPDC1, KRAS, p-ERK1/2 and ERK1/2 protein expression. Glucose uptake and lactic acid detection were also used to detected glycolysis in Huh-7 cells after transfected DEPDC1 siRNA and NC. The DEPDC1 downregulation-induced KRAS inhibition was restored by KRAS expression plasmid, and the role of KRAS in DEPDC1 regulated glycolysis, proliferation and migration of Huh-7 cells was detected by glucose uptake kit, lactate detection kit, CCK-8 and Transwell chamber. 21 038 genes were changed in DEPDC1 downregulated Huh-7 cells compared with control, wherein, a total of 9 781 genes were upregulated and 11 257 genes downregulated. GSEA results showed that the differentially expressed genes in DEPDC1 downregulated Huh-7 cells were significantly enriched in the central carbon metabolism signaling pathway. DEPDC1 knockdown significantly decreased the glucose uptake, lactate production, and protein levels of KRAS and p-ERK1/2. However, KRAS overexpression significantly reversed the inhibition of glucose uptake, lactate production, proliferation and migration induced by DEPDC1.In total, these results suggest that DEPDC1 may regulate glycolysis through KRAS/p-ERK1/2 pathway, theraby affect HCC progression