This paper investigates the effect and mechanism of nodakenin (NK) on hepatocellular carcinoma cells.HepG2 and MHCC-97H cells were treated with different concentrations of NK.Cell proliferations were detected by CCK-8,and migration and invasion abilities were observed by cell scratch and Trans-wells experiments.NK significantly inhibited the proliferation,migration and invasion of HepG2 and MHCC-97H cells in vitro.RNA-seq was conducted to analyze the expression of each gene before and after the administration of NK.qRT-PCR,Western blotting and cellular immunofluorescence were used to verify the RNA-seq result.The results showed that the expression of chemokine 5 (CXCL5) was significantly decreased after administration of NK,and the phosphorylation of ERK and MEK was inhibited at the same time.In summary,it can be concluded that NK inhibits ERK/MEK signaling pathway by down-regulating CXCL5,thereby inhibiting hepatocellular carcinoma.