We verified via qPCR and WB experiments that the expression of TM4SF1 was positively correlated with the expression of the NOTCH pathway ligand JAGGED1, receptor NOTCH1, and target gene HES1 in HCC cells, meanwhile, with the help of the NOTCH pathway targeted inhibitor RO4929097, we found that overexpression of TM4SF1 could reverse the inhibitory effect of RO4929097 on the NOTCH pathway. The gene discussed is NOTCH1; the disease is hepatocellular carcinoma.