Effects Of IL-33/ST2 On Proliferation,Apoptosis,Migration And Invasion Of Human Liver Cancer Cells

Objective: To explore the effects of IL-33 and its receptor ST2 on the proliferation,the apoptosis,migration and invasion of huh-7 and Hep3 B human hepatocellular carcinoma cells were investigated in vitro,it is to provide new ideas for the basic and clinical research of hepatocellular carcinoma.Methods:(1)First,MTT assay was used to detect the cell survival of huh-7 and Hep3 B human liver cancer cells with different concentrations of IL-33: the concentrations were divided into 0ng/L,20ng/L,40ng/L,and 60ng/L,and the absorbance value(OD value)of each group was detected.Next,blank control group,IL-33 group,si-ST2 group and IL-33 +si-ST2 group were set.(2)The effect of IL-33 and si-ST2 on the survival rate of huh-7 and Hep3 B human liver cancer cells was detected by MTT assay,and the absorbance value(OD value)of each group was detected.(3)Flow cytometry was used to detect the effects of IL-33 and si-ST2 on the apoptosis of huh-7 and Hep3 B human hepatocellular carcinoma cells..(4)The influence of IL-33 and si-ST2 on huh-7 and Hep3 B human liver cancer cell migration was detected by scratch test.The width and mobility of scratches in different groups at different times [(0h cell spacing-24 h cell spacing)/0h cell spacing *100%] were measured in the AI software.(5)Transwell invasion assay was used to detect the effects of il-33 and si-ST2 on the invasion of huh-7 and Hep3 B human liver cancer cells,and the cells in each group that entered the lower compartment membrane were detected.Two independent sample t test was used for comparison between the two groups,one-way analysis of variance was used for comparison between the two groups,and Spearman rank correlation was used for correlation analysis.P < 0.05 indicated that the difference was statistically significant.Results:(1)In the MTT assay after the action of different concentrations of IL-33 on huh-7 and Hep3 B human liver cancer cells,the higher the concentration of IL-33,the greater the OD value,and the more living cells,the higher the survival rate of huh-7 and Hep3 B human liver cancer cells.After Hep3 B and huh-7 cells were treated with IL-33 at different concentrations in the four groups,their OD values were statistically significant(P <0.05).In Hep3 B cells,OD values of 20ng/L,40ng/L and 60ng/L were compared with those of0ng/L,and OD values of 20ng/L were statistically significant compared with those of 60ng/L(P < 0.05).There was no statistically significant difference between the OD value of 20ng/L and that of 40ng/L,and that of 40ng/L and that of 60ng/L(P > 0.05).In huh-7 cells,OD values of 40ng/L and 60ng/L were compared with those of 0ng/L,OD values of 20ng/L were compared with those of 60ng/L,OD values of 40ng/L were statistically significant(P <0.05).The OD values of 20ng/L were not statistically significant compared with those of0ng/L and 40ng/L(P > 0.05).The analysis of the correlation between IL-33 concentration and OD value showed that the cell survival rate showed a significant positive correlation with IL-33 concentration(P < 0.05).(2)for huh-7 and Hep3 B human hepatocellular carcinoma cells MTT after transfection with si-ST2,the cell survival rate of the IL-33 treatment group was significantly higher than that of the control group,and that of the si-ST2 treatment group was significantly lower than that of the control group.The cell survival rate of the IL-33 +si-ST2 group was significantly higher than that of the si-ST2 group,and the cell survival rate of the IL-33 + si-ST2 group was significantly lower than that of the IL-33 group(P <0.01).(3)for the modulation experiment of huh-7 and Hep3 B human hepatocellular carcinoma cells after transfection with si-ST2,the apoptosis rate of the IL-33 treatment group was lower than that of the control group.The apoptosis rate of si-ST2 group was higher than that of control group and IL-33 treatment group.The apoptosis rate of the IL-33 + si-ST2 group was lower than that of the si-ST2 group,and the apoptosis rate of the IL-33 + si-ST2 group was higher than that of the IL-33 group.Conclusion: IL-33 may be involved in the proliferation,apoptosis,migration and invasion of huh-7 and Hep3 B hepatocellular carcinoma cells through the ST2 pathway.