The Effect Of Human Embryonic Stem Cells On Acute Myeloid Leukemia Cells

ObjectiveTo determine the effect of embryonic stem cell on the proliferation and apoptosis in human acute myeloid leukemia cell line KG1 a and explore the potential mechanism.MethodsEstablish the direct co-culture system between human embryonic stem cell line H9 and human acute myeloid leukemia cell line KG1 a in vitro.The proliferation of suspension cell in co-culture system was tested by CCK8 assay.The cell cycle was analyzed using flow cytometry(FCM).The expressions of P21,Cyclin-D1,CDK4 in KG1 a cells were detected with the use of real time PCR(RT-PCR)and Western blot.The apoptosis was detected by FCM.The expression of Bax,Caspase-3 and Bcl-2 in suspension cell in co-culture system were detected by RT-PCR and Western Blot.The changes of PTEN / PI3 K / AKT signaling pathway in suspension cell in co-culture system were detected by RT-PCR and Western Blot.ResultsAfter co-cultured with H9 cells,the proliferation of suspension cell in co-culture system was significantly inhibited and the cell cycle was arrested at the G2/M phase;the mRNA expression of the cycle-related genes was detected by fluorescent quantitative PCR and found that the mRNA expression of Cyclin-D1 and CDK4 was down-regulated,and P21 expression was up-regulated.Western Blot showed that the protein expression was consistent with that of PCR,and the difference was statistically significant(P<0.05).After co-cultured with H9 cells,the apoptosis of suspension cell in co-culture system increased significantly.Annexin V-FITC/PI double staining flow cytometry showed that the apoptosis rate of suspension cells in co-culture system was higher than that of the control group.Bax and Caspase-3 mRNA expression of suspension cell in co-culture system was up-regulated and Bcl-2 mRNA expression was down-regulated.Western Blot showed that the protein expression was consistent with that of PCR and the difference was statistically significant(P<0.05).After co-culture with H9 cells,the mRNA and protein contents of signal molecules in the PI3K/AKT signaling pathway of suspension cells in co-culture system also changed.The m RNA and protein content of PTEN genes were up-regulated,and the mRNA and protein content of total AKT remained unchanged.The amount of phosphorylated AKT(pAKT)was reduced.ConclusionsEmbryonic stem cell can inhibit the proliferation of KG-1a and induce the apoptosis,which might be related to the activation of PTEN / PI3 K / AKT signaling pathway...