Sensitivity Screening Of Small Molecule Kinase Inhibitors And Research On Anti-hepatocarcinoma Of WH-4-023

Objective:To investigate the effects of SRC kinase inhibitor WH-4-023 on proliferation and apoptosis of SNU-423 and BEL-7402 and their mechanisms.Methods:1.MTT assay,plate clone formation test,flow cytometry and Hoechet staining were used to detect the effects of inhibitors on the cell proliferation and apoptosis,the cells were treated by the following drugs concentrations:WH-4-023（0μmol/L、0.2μmol/L、1μmol/L、2μmol/L）on hepatocellular carcinoma cell line SNU-423,WH-4-023（0μmol/L、0.3μmol/L、1.5μmol/L、3μmol/L）on hepatocellular carcinoma cell line BEL-7402.2.Western blot was used to detect the effects of WH-4-023 on the expression of SRC/STAT3 pathway-related proteins and anti-apoptotic protein Bcl-2.Results:1.MTT results showed that the IC₅₀0 of SRC kinase inhibitor WH-4-023 on hepatocellular carcinoma cell line SNU-423 and hepatocellular carcinoma cell line BEL-7402 were 1.15μmol/L and 1.456μmol/L,respectively.2.MTT results showed that the survival rate of hepatocellular carcinoma cell line SNU-423 decreased with the increase of drug dosage after treatment with different concentrations of WH-4-023.Plate clone formation experiment showed that the number of clone formation of SNU-423 decreased in a dose-dependent manner.Flow cytometry results showed that there was no significant difference in cell percentage between the control group and the drug group at each stage.Western blot results showed that WH-4-023 could inhibit the expression of p-SRC,p-STAT3 and Bcl-2 in a dose-dependent manner at low concentration,but had no effect on the expression level of SRC and STAT3 proteins.3.MTT results showed that the survival rate of hepatocellular carcinoma cell line BEL-7402 decreased with the increase of drug dosage after treatment with different concentrations of WH-4-023.Plate clone formation experiment showed that the number of clone formation of BEL-7402 decreased in a dose-dependent manner.Flow cytometry results showed that there was no significant difference in cell percentage between the control group and the drug group at each stage.Western blot results showed that WH-4-023 could inhibit the expression of p-SRC,p-STAT3 and Bcl-2 in a dose-dependent manner at low concentration,but had no effect on the expression level of SRC and STAT3 proteins.Conclusions:1.WH-4-023 has strong anti SNU-423 ability and strong anti BEL-7402 ability.At low concentration,WH-4-023 can inhibit the proliferation of two kinds of hepatocarcinoma cells and induce apoptosis.2.Preliminary studies have shown that WH-4-023 may inhibit the proliferation of hepatocellular carcinoma cell line SNU-423 and hepatocellular carcinoma cell line BEL-7402 and induce apoptosis of the two kinds of hepatocellular carcinoma cells by inhibiting the phosphorylation of SRC/STAT3 pathway-related proteins and the expression of Bcl-2.