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.