STYK1/NOK Effects Of HepG2 And Huh-7 Cells On Apoptosis,Proliferation And Cell Cycle

Objective: To investigate the effect of STYK1/NOK gene on HepG2 and huh-7 in hepatocellular carcinoma cell lines.To investigate the effect of STYK1/NOK overexpression on apoptosis and cell proliferation of HepG2 cells,and to reverse verify the effect of STYK1/NOK protein expression inhibition on apoptosis,proliferation and cell cycle of HepG2 cells.Subsequently,it was verified in huh-7cells.Methods: STYK1/NOK was overexpressed in HepG2 cells,and the expression of STYK1/NOK was determined by western blotting.HepG2 cells transfected with 0,2,6 μg STYK1/NOK plasmids were respectively transfected with Annexin V-FITC/PI apoptosis assay kit to detect the apoptosis of HepG2 cells.After transfection with 0,2,6 μg STYK1/NOK plasmids,the effect of overexpression of NOK on HepG2 cell proliferation was detected by CCK8 cell proliferation assay 48 hours later.In order to determine the test results,siRNA was used to inhibit the expression of STYK1/NOK in HepG2 cells,and cell proliferation,apoptosis and cell cycle experiments were conducted.Western blotting was used to detect the expression of apoptosis-related proteins caspase 3 and cleaved-caspase 3 to determine the effect of STYK1/NOK on the apoptosis of HepG2 cells.Subsequently,the specific action mechanism of STYK1/NOK on HepG2 of liver cancer cells was determined.Subsequently,experiments were performed on huh-7 cells.Results: The results showed that the expression of STYK1/NOK was correlated with the transfected plasmid concentration,showing a dose-dependent effect.The study found cell apoptosis percentage gradually decrease in transfection 0,2,6 μg STYK1 /NOK plasmid group.Repeating three groups,using t-test statistics,statistical significance(P<0.05),suggesting that the transfection STYK1/NOK inhibits apoptosis of HepG2 cells.CCK8 cell proliferation assay was used to investigate the effect of STYK1/NOK gene expression on cell proliferation.48 h after transfection,CCK8 reagent was added and subsequently detected,and it was found that STYK1/NOK overexpression promoted HepG2 cell proliferation.Subsequently,the apoptotic protein caspase 3,cleaved-caspase 3 was detected.In this study,we foundthat the expression of caspase 3 did not decrease and the expression of cleavedcaspase 3 did not increase after the overexpression of STYK1/NOK,indicating that the overexpression of STYK1/NOK did not activate the apoptosis of HepG2 cells.To further verify the test results,siRNA was used to inhibit the expression of STYK1/NOK in HepG2 cells.The results showed that the inhibition of STYK1/NOK protein expression promoted the cell apoptosis,and the apoptosis rates were 30.2%,30.6% and 36.5%,respectively,after transfection of 0,2,6 μg si-STYK1 /NOK plasmids.There was a statistically significant difference in the apoptosis rate between the 0 μg si-STYK1 /NOK plasmid group and the 6μg si-STYK1 /NOK plasmid group(P<0.05).Cell proliferation assay showed that STYK1/NOK inhibited HepG2 cell proliferation(P<0.01).The inhibition of STYK1/NOK protein expression resulted in decreased caspase 3 and increased cleaved-caspase 3.Cell cycle results showed that after inhibiting the expression of STYK1/NOK protein,cells in S phase increased and cells in G0/G1 and G2/M phases decreased,indicating that STYK1/NOK inhibition blocked HepG2 cells in S phase,thus inhibiting cell proliferation.Subsequently,the results of apoptosis experiment and cell proliferation experiment were consistent with HepG2 results in huh-7 cells.After inhibiting the expression of STYK1/NOK protein in huh-7 cells,apoptosis was promoted by activation of caspase 3.Conclusion: Studies have shown that STYK1/NOK inhibition can promote apoptosis of HepG2 cells and inhibit cell proliferation by targeting caspase 3.Cell cycle detection results showed that cells in S phase increased significantly,indicating that STYK1/NOK inhibition blocked cells in S phase,inhibited cell cycle progression and inhibited cell proliferation in HepG2.On the contrary,the results showed that STYK1/NOK could promote cell proliferation and inhibit apoptosis.The experimental results showed that STYK1/NOK might be a target gene for the treatment of liver cancer.However,this study is mainly conducted at the cellular level,further validation by in vivo experiments is needed.