Objective: To investigate the effect of prostaglandin E2 recoptor 4 antagonist (EP4A) on the self-renewal ability of human CD34+ cells and its mechamism.
Methods: The peripheral blood hematopoietic stem cell of 20 healthy donors received the G-CSF-mobilization were collected, then the human CD34+ cells were sorted out by MACS microbead kit. The human CD34+ cells were treated with DMSO (control group), EP4A (EP4A group) and EP4A+EP4A antagonist (EP4A+EP4A group) for 72 hours. The differential genes and pathways related with CD34+ cell stemness were detected by Thermogram and Pathway enrichment analysis. and then the expression levels of protein and gene (β-catenin, Nanog, Oct4, Sox2, Stat3, AKT, P38) were detected by qRT-PCR and Western blot respectively.
Results: EP4A could elevate the mRNA and protein expression of β-catenin, Nanog, Oct4, Sox2, in comparison with control group, however, mRNA and protein expression of STAT3, AKT, P38 were not changed. When human CD34+ cell were cultured with EP4A+XAV939 it was found that the mRNA and protein expression of β-catenin was downregulated, moreover the mRNA and protein expression of Nanog, Oct4, Sox2 were reduced.
Conclusion: EP4A can upregulate stemness factors-β-catenin, Nanog, Oct4 and Sox2 in human CD34+ cell in vitro, but not STAT3, AKT and P38.
题目: EP4A对人CD34+细胞干性因子表达的影响.
目的: 探讨前列腺素E2特异性受体激动剂4（EP4A）增强人CD34+细胞干性因子表达的机制.
方法: 收集中山大学附属第一医院血液科20例健康供者经G-CSF动员后的外周血造血干细胞采集物，采用免疫磁珠法分选出人CD34+细胞。以DMSO作对照，分别用EP4A、EP4A + EP4A拮抗剂（EP4AA）作用人CD34+细胞72 h后，应用热图和Pathway富集分析检测与CD34+细胞干性相关的差异基因和通路；再用qRT-PCR和Western blot检测不同组别细胞中通路蛋白和差异基因的mRNA及蛋白（β-catenin、Nanog、Oct4、Sox2、Stat3、AKT、P38）表达水平.
结果: 与对照组相比，EP4A组人CD34+细胞中β-catenin、Nanog、Oct4及Sox2 mRNA及其蛋白的表达均增高，而STAT3、AKT、P38 mRNA及其蛋白的表达无变化； EP4A与EP4AA（XAV939）共同作用人CD34+细胞时，人CD34+细胞β-catenin表达水平明显降低，并且Nanog、Oct4及Sox2基因及蛋白表达相应减少.
结论: EP4A可增加人CD34+细胞干性因子（β-catenin、Nanog、Oct4和Sox2）表达，而不能增加STAT3、AKT、P38表达.