The phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway is abnormally activated in hepatocellular carcinoma and promotes the occurrence and development of cancer. In hepatocellular carcinoma, the heat shock protein Hsp70 binding protein 21（Hsp70 binding protein 21） is in a low expression state, and overexpression of HBP21 significantly induces cancer cell apoptosis. Using qRT-PCR technology to detect the mRNA level of HBP21 in liver cancer tissues, it is found that the mRNA level of HBP21 in cancer tissues is lower than that in adjacent tissues. Hepatocellular carcinoma cell Huh7 is treated with insulin to activate the PI3K/AKT signaling pathway in the cells, and the transcription and translation levels of HBP21 in the cells are detected by qRT-PCR technology and Western blotting. With the prolonged insulin treatment time, the mRNA and protein levels of HBP21 show a downward trend. Overexpression of HBP21 in Huh7 cells significantly inhibits insulin-induced phosphorylation of AKT and mammalian target of rapamycin（mTOR）；through ubiquitination and western blotting experiments, it is found that HBP21 does not affect AKT K48 and K63 ubiquitination as well as phosphatase and tensin homolog deleted on chromosome ten（PTEN） protein levels. The inhibitor LY294002 was used to treat Huh7 cells to confirm that HBP21 acts on the PI3K/AKT signaling pathway. Further research shows that HBP21 does not affect the mRNA levels of IRS1 and IRS2, but inhibits the phosphorylation of insulin receptor beta（IRβ） and affects the activation of the PI3K/AKT signaling pathway. In Huh7 cells，HBP21 plays an important role in inhibiting abnormal activation of PI3K/AKT in liver cancer cells. The lack of expression of HBP21 in hepatocellular carcinoma and its inhibition of PI3K/AKT signaling pathway make it possible to become a potential target for cancer treatment.