Objective To investigate the effects and molecular mechanism of EEATK on proliferation, apoptosis of human hepatocellular carcinoma Hep3B cells and Huh-7 cells and to explore its underlying mechanism . Methods Human hepatoma cells Hep3B cells and Huh-7 cells cultured in vitro, blank control group, Sorafenib group (5uM), EEATK groups (0.10 mg/mL、0.15 mg/mL、0.20 mg/mL、0.3 mg/mL)were set and given corresponding drug intervention, respectively. CCK-8 assay was used to measure the viability of different interventions on the proliferation of Hep3B cells and Huh-7 cells, to screen the optimal action concentration for subsequent experiments. Hep3B cells and Huh-7 cells were divided into the blank control group and EEATK group(0.15mg/mL) treated with for 24 hours,EdU staining assay, colony formation assay were used to explore the effect of EEATK on the proliferation , the apoptosis rate,were detected by Annexin V-FITC/PI double staining assay.The transcriptome sequencing(RNA-seq)technology was used to analyze differentially expressed genes (DEGs) related to cell proliferation, apoptosis on Hep3B cells in blank control group and EEATK group(0.15mg/mL)of Hep3B cells, DEGs were analyzed for Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. CTD (Comparative Toxicogenomics Database) was used to validate the expression of key proteins related to cell proliferation, apoptosis. Results Compared with the blank control group, different concentration of EEATK could significantly inhibit the activity of Hep3B and Huh-7 cells (P <0. 01) ; After treated with 0.15 mg/mL EEATK, the EdU positive cell rate and cell clone formation rate were significantly decreased (all P <0. 01), at the same time ,the apoptotic rates of EEATK group were significantly increased(P < 0.01). Compared with the blank control group, The transcriptome sequencing of Hep3B cells showed that EEATK induced significant changes in the expression of 1577 DEGs (P <0. 01) , of which 942 were up-regulated and 635 were down-regulated. GO functional enrichment analysis revealed that DEGs were mainly enriched in cholesterol synthesis, inflammation, and extracellular matrix. KEGG pathway analysis showed that EEATK may mainly pass through TFG- β Signal pathways and NF-κB signaling pathway plays an anti-tumor role.DEGs enrichment analysis showed that EEATK can significantly downregulate the expression of apoptosis related genes such as BIRC5 and CDK1 (P<0.01), the expression of CDKN1A and EGLN3 were significantly upregulated (P<0.01), At the same time, EEATK can cause significantly downregulation of cell proliferation related genes such as FAM83D and Ki-67 (P<0.01), mean while, the expression of MYC and FoxC1 were significantly upregulated (P<0. 01).