Further by in vitro cellular experiments, it was found that after incubation with isoscopoletin, the relative cellular activity of HepG2 cells decreased, and intracellular glucose consumption and lactate production decreased, along with decreased levels of glycolysis-related genes and proteins, GPD2, GPI, Hsp90α, and PGK2, suggesting that isoscopoletin inhibits the glycolysis process by modulating glycolysis-related proteins, thereby inhibiting HCC cell proliferation, which verified the results of network pharmacology and MST experiments. This evidence concerns the gene GPD2 and hepatocellular carcinoma.