The Influence Of ATF4 To PKM2 And Hepatocellular Carcinoma Cell HepG2 Apoptosis

Objective: Study the influence of activation of transcription factor 4(ATF4)overexpression to hepatocellular carcinoma cell HepG2 apoptosis and ATF4 down-regulated the expression of the Warburg effect protein PKM2.Methods:In this experiment,the hepatocellular carcinoma cell HepG2 has been divided into three groups: blank control group;the empty plasmid transfection group;transfection ATF4 plasmid.Build ATF4 plasmid and transfect cell HepG2 with the method of instantaneous transfection by liposome mediate.Observe the transfection rate under the fluorescence microscope.Western-Blot detect the expression of ATF4 protein.Lactic acid detection kit detect the lactic acid contents of nutrient solution.Western-Blot detect the expression of PKM2 protein.To detect cell apoptosis rate using flow cytometry.Results: 1.According to the results of Transient transfection,The figures of HepG2 cells in the fluorescence microscope visible horizon showed most visible green fluorescent,whose transfection rate was 75%.2.Western-Blot showed that compared with blank control group and the empty plasmid transfection group,the expression of ATF4 protein increased significantly(P < 0.05),and the expression of blank control group and group of transfection ATF4 plasmid were no statistically significant difference(P≥0.05).3.The results of Lactic acid content detecting showed that compared with blank control group(10.17±1.25)mmol/L and the empty plasmid transfection group(10.21±1.53)mmol/L,group of transfection ATF4 plasmid HepG2 cells lactic acid content(5.63±1.05)mmol/L significantly reduced(P < 0.05),and the lactic acid content of blank control group and group of transfection ATF4 plasmid were no statistically significant difference(P≥0.05).4.Western-Blot showed that compared with blank control group and the empty plasmid transfection group,the expression of PKM2 protein reduced significantly(P < 0.05)in HepG2 cells of group of transfection ATF4 plasmid,and the expression of blank control group and group of transfection ATF4 plasmid were no statistically significant difference(P≥0.05).5.Flow cytometry instrument testing results show that compared with blank control group(17.62±0.83%)%and the empty plasmid transfection group(17.80±0.84)%,group of transfection ATF4 plasmid HepG2 cells ’ apoptosis rate(43.76±5.61)%increased significantly(P < 0.05),and apoptosis rate of blank control group and group of transfection ATF4 plasmid were no statistically significant difference(P≥0.05).Conclusion:.ATF4 overexpression promoted the hepatocellular carcinoma cell HepG2 apoptosis and inhibited its Warburg effect through down-regulated the expression of Warburg effect-related protein PKM2.